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  • 51.
    Källman, Jan
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Group B streptococcal infections in neonates: Clinical and pathogenic aspects1997Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Clinical and pathogenic aspects of Group B streptococci (GBS), as a major pathogen responsible of invasive disease in newborn infants, were investigated.

    Cases of neonatal septicaemia during 1981-1994 were studied at Orebro Medical Centre Hospital. 132 children ful1filled laboratory and clinical criteria for neonatal septicaemia. The annual incidence increased significantly, from 2.3 cases during the first 7-year period to 3.3 per 1000 live births during the second 7-year period. The increase in incidence between the two 7- year periods was almost entirely due to an increase in Staphylococcus aureus ( from 9 to 32, p<O.Ol) and coagulase-negative staphylococci (CoNS) (from 7 to 20, p<0.05) and mainly affected preterm neonates 48h or more after delivery while GBS infection usually occurred in full-term children during the first 48h of life. An increased resistance among CoNS to methicillin and gentamicin was observed between the first and second 7-year period.

    To study the ability of GBS to adhere to the target cell, a cell-culture model with human umbilical vein endothelial cells (HUVEC) was used. Clinical isolates of serotype Ill adhered significantly (p=O.OOOl) better than other serotypes. Isogenic variants of serotype Ill with low amounts of capsule substance adhered significantly (p=O.OOl) better to the HUVEC than variants expressing high amount of capsule substance. The role of GBS capsular substance as a major virulence factor is further underlined by the fact that it impair the phagocytic capacity by polymorphonuclear leukocytes (PMNL). Growth conditions for GBS, simulating different in vivo environments, greatly affect capsule expression.

    The extent of and the penetration route of GBS over epithelial linings was examined with a model using Madin Darby Canine Kidney cells (MDCK). It was demonstrated that GBS can penetrate intact polarized MDCK cells by transcytosis in a selective apical-to-basolateral direction and the mechanism for this is metabolically dependent.

    In a chemiluminescence assay (CL), PlviNL function and opsonic capacity were shown to be significantly impaired in neonates and correlate to maturation of the newborn child. This combined defect in cellular and humoral defences may contribute to the increased susceptibility to GBS infection in preterm infants.

    In a prospective study in newborn having suspected sepsis, IL-6 and C-reactive protein (CRP) were measured early. Early-sampled IL-6 levels were significantly better than early-sampled CRP levels in distinguishing between mild respiratory disorders and septicaemia in the newbom child.

  • 52.
    Kühme, Tobias
    et al.
    Malmö University Hospital.
    Isaksson, Barbro
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Dahlin, Lars-Göran
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för medicin och vård, Thoraxkirurgi. Östergötlands Läns Landsting, Hjärtcentrum, Thorax-kärlkliniken.
    Wound contamination in cardiac surgery, a systematic quantitative and qualitative study of bacterial growth in sternal wounds in cardiac surgery patients2007Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 115, s. 1001-1007Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Objectives: To investigate the degree of bacterial contamination in the sternal wound during cardiac surgery and the sternal skin flora after operation in order to increase our understanding of the pathogenesis of sternal wound infections. Design: Prospective study where cultures were taken peri- and postoperatively from sternal wounds and skin. Setting: University Hospital. Patients: 201 cardiac surgery patients. Results: 89% of the patients grew bacteria from the subcutaneous sternal tissue. 98% of the patients showed bacterial growth on the surrounding skin at the end of the operation. We found both commensal and nosocomial bacteria in the sternal wound. These bacteria had different temporal distribution patterns. Coagulase-negative staphylococci (CoNS) and Propionibacterium acnes (PA) were by far the most prevalent bacteria during and after the operation. Furthermore, 41% of patients had more than 10 000 CFU/pad CoNS on the skin. There was no correlation between length of operation and number of bacteria. Men displayed higher bacterial counts than women on the skin. Conclusion: Skin preparation with ethanol/chlorhexidine is unable to suppress the physiological skin flora for the duration of a heart operation. A decrease of CoNS and PA postoperatively can be caused by competitive recolonisation of commensal and nosocomial bacteria.

  • 53.
    Larsson, Per-Göran
    et al.
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Genus och medicin.
    Fåhraeus, Lars
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Obstetrik och gynekologi. Östergötlands Läns Landsting, Barn- och kvinnocentrum, Kvinnokliniken i Linköping.
    Carlsson, Bodil
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Jakobsson, Tell
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Forsum, Urban
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Late miscarriage and preterm birth after treatment with clindamycin: A randomised consent design study according to Zelen2006Ingår i: British Journal of Obstetrics and Gynecology, ISSN 1470-0328, E-ISSN 1471-0528, Vol. 113, nr 6, s. 629-637Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Objective: To screen for bacterial vaginosis (BV) and to investigate the effect of treatment with vaginal clindamycin in order to observe the effect on late miscarriage and delivery prior to 37 completed weeks (primary outcome). Design: Randomised consent design for clinical trials according to Zelen. Setting: Southeast region of Sweden. Population: A total of 9025 women were screened in early pregnancy. Methods: A total of 819 women with a Nugent score of 6 and above were considered to have BV and treated according to Zelen allocation. The incidence of late miscarriage and spontaneous (noniatrogenic) preterm birth was assessed. Main outcome measures: Late miscarriage and spontaneous preterm delivery before 37 weeks. Results: Therapy with vaginal clindamycin had no significant impact on the incidence of spontaneous preterm delivery prior to 37 completed weeks, OR 0.90, 95% CI 0.40-2.02 (primary outcome variable). However, only 1 of 11 women in the treatment group versus 5 of 12 in the control group delivered prior to 33 completed weeks, OR 0.14, 95% CI 0.02-0.95. Treatment was associated with 32 days longer gestation for the 23 participants who had late miscarriage or spontaneous preterm birth (P= 0.024, Mann-Whitney U test) and significantly fewer infants had a birthweight below 2500 g (secondary outcome). A follow up of infants born preterm 4 years postnatally indicated that extending gestational age did not increase the number of sequelae. Conclusions: Clindamycin vaginal cream therapy was associated with significantly prolonged gestation and reduced cost of neonatal care in women with BV. Early screening for BV and treatment with clindamycin saved approximately €27 per woman. © RCOG 2006 BJOG An International Journal of Obstetrics and Gynaecology.

  • 54. Larsson, PG
    et al.
    Forsum, Urban
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Diagnostik. Diagnostikmetoder2004Ingår i: Bakteriell Vaginos / [ed] Mats Bergström;P-G Larsson,Urban Frosum och 3M Pharma., Växjö: Grafiska Punkten , 2004, s. 49-57Kapitel i bok, del av antologi (Övrigt vetenskapligt)
  • 55.
    Larsson, P-G
    et al.
    Avd för Obstretik och Gynekologi Kärnsjukhuset, Skövde.
    Fåhraeus, Lars
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Obstetrik och gynekologi. Östergötlands Läns Landsting, Barn- och kvinnocentrum, Kvinnokliniken i Linköping.
    Carlsson, Bodil
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Jakobsson, Tell
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Obstetrik och gynekologi.
    Forsum, Urban
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Predisposing factors for bacterial vaginosis, treatment efficacy and pregnancy outcome among term deliveries, results from a preterm delivery study2007Ingår i: BMC Women's Health, E-ISSN 1472-6874, Vol. 7Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Background: Bacterial vaginosis (BV) during pregnancy is associated with an increased risk of preterm delivery but little is known about factors that could predict BV. We have analyzed if it is possible to identify a category of pregnant women that should be screened for BV, and if BV would alter the pregnancy outcome at term, we have also studied the treatment efficacy of clindamycin. Methods: Prospective BV screening and treatment study of 9025 women in a geographically defined region in southeast Sweden. BV was defined as a modified Nugent score of 6 and above. Data was collected from the Swedish Medical Birth Register. Women allocated to treatment were supplied with vaginal clindamycin cream. The main outcome goals were to identify factors that could predict BV. Results: Vaginal smears were consistent with BV criteria in 9.3%. Logistic regression indicates a significant correlation between smoking and BV (p < 0.001) and a greater prevalence of BV in the lower age groups (p < 0.001). We found no correlation between BV and history of preterm deliveries, previous miscarriages, extra-uterine pregnancies, infertility problems or reported history of urinary tract infections-factors that earlier have been associated with BV. Treatment with clindamycin cream showed a cure rate of 77%. Less than 1% of women with a normal vaginal smear in early pregnancy will develop BV during the pregnancy. There was no association between BV and the obstetric outcome among women who delivered at term. Women with BV, both treated patients and nontreated, had the same obstetric outcome at term as women with normal vaginal flora. Conclusion: BV is more than twice as common among smokers, and there is a higher prevalence in the younger age group. However these two markers for BV do not suffice as a tool for screening, and considering the lack of other risk factors associated with BV, screening of all pregnant women might be a strategy to follow in a program intended to reduce the number of preterm births. © 2007 Larsson et al, licensee BioMed Central Ltd.

  • 56.
    Liss, Per-Erik
    et al.
    Linköpings universitet, Institutionen för hälsa och samhälle.
    Aspevall, Olle
    Karolinska institutet.
    Karlsson, Daniel
    Linköpings universitet, Tekniska högskolan. Linköpings universitet, Institutionen för medicinsk teknik, Medicinsk informatik.
    Forsum, Urban
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Interpreting definitions: The problem of interpreting definitions of medical concepts2004Ingår i: Medicine, Health care and Philosophy, ISSN 1386-7423, E-ISSN 1572-8633, Vol. 7, s. 137-141Artikel i tidskrift (Refereegranskat)
  • 57.
    Lymer, Ulla-Britt
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Blood exposure in health care: health care workers' and patients' experiences2004Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    The general aim of this thesis was to describe and analyse health care workers' blood-exposure incidents, protective measures, and the motives behind decision making about risks and protective measures. The aim was also to procure knowledge about patients' conceptions about their care, and if precautions taken by the health care workers were included in quaiity care. The methods used were: in study I quantitative method, in studies II and III grounded theory method, and in study IV phenomenographic method.

    The aims of study I were to describe and analyse blood-exposure incidents and compliance in relation to recommended serological investigations, universal precautions and incident reporting routines and also to follow up a campaign against blood-exposure. Instruments used were incident report forms (n=473) and questionnaires (n=132), (n=108), (n=517). The majority of the 473 reported blood-exposure incidents came from nurses and the minority from physicians. Incidents occurred most often on hospital wards, the most commonly reported incident being needle-stick injuries, 35% ofwhich occurred during recapping the needle. Medicallaboratory technicians reported significantIy more mucocutaneous incidents than other professionaIs (P<0,01). Serological investigations post-exposure varied and 35% of individuaIs were not tested. In an anonymous questionnaire, the respondents recalled 1180 incidents, although onIy 9% of these were reported. Physicians reported onIy 3% of these, medical laboratory technicians reporting 36%. The under reporting was most common in operation theatres and in anaesthesia. Eighty-one percent believed that the accident could have been avoided if they had followed the recommended clinical practice. Despite knowledge of universal precautions, professionaIs continue to behave in a risky manner, which can result in blood exposure incidents and possibly a blood-borne viral infection.

    The aim of study II was to identify factors that influenced health care workers' actions in situations involving a risk of blood-exposure. Nurses and assistant nurses were interviewed (n=15). The analysis showed that there was a conflict between different demands involving protecting the patient's privacy, protecting themselves from being infected and respecting the norms of the department. The process of managing this conflict was labelled 'balancing', which most often resulted in the choice of a diagnosis-related strategy, i.e. a non-compliant behaviour. The underlying causes of how patterns of action are formed by individual and socio-cultural forces resulted in five categories, which were seen as forces that could underrnine compliance.

    The aim of study III was to describe and analyse different forces that promote adherence to universal precautions. Nurses and assistant nurses were interviewed (n=15, the same as in study II). The charge nurse, informal leaders, students, infection controi nurses, type of work., availability of equipment, blood-exposure incidents and media-coverage of viral blood-borne infections were described as potentially irnportant for compliance. The properties these agents must possess in order to be influential were also described. The results irnply that information about safe practices alone is insufficient to achieve that goal. All factors of importance for compliance must be taken into consideration in clinical work and in education.

    The aim in study IV was to identify and describe patients' conceptions of quality care and of barrier care. The patients (n=14) were adult and treated for orthopaedic reasons. Included in their conceptions of quality care were: Nice manners, mutual achievement, being involved, being cured, being cared for, and having safe care. When comparing these conceptions with previous research about patients' views of quaIity care, the findings confirmed, to a large extent, the findings from other studies. Included in patients' conceptions of barrier care were: Regular use of gloves, regular use of masks and eye-shields, use of gloves in special situations, use of masks and eye-shields in special situations, and keeping clean. The conceptions were of an interpersonal, as well as of a medical-technical nature. Patients' conceptions of barrier care could be included in the category: Having safe care. The frequentIy expressed opinion, among nurses and assistant nurses, that patients may be offended by the use of protective equipment could be refuted.

    This thesis has contributed to an improved understanding of the occurrence and handling of blood-exposure incidents. The dynamics of compliance and non-compliance with universal precautions have also been described by means of an attempt to uncover the interplay between deactivators and re-activators in the safety culture on wards. Patients' conceptions about barrier care were shown to be an integrated part of quality care.

    Delarbeten
    1. A descriptive study of blood exposure incidents among healthcare workers in a university hospital in Sweden
    Öppna denna publikation i ny flik eller fönster >>A descriptive study of blood exposure incidents among healthcare workers in a university hospital in Sweden
    1997 (Engelska)Ingår i: Journal of Hospital Infection, ISSN 0195-6701, E-ISSN 1532-2939, Vol. 35, nr 3, s. 223-235Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    In an attempt to document blood exposure incidents and compliance with recommended serological investigations, universal precautions and incident reporting routines, data was collected from occupational injury reports during a two-year period. In addition, a sample of healthcare workers (HCWs) answered a questionnaire about blood tests and work routines. In a third part of the study some HCWs were asked about the type and actual frequency of incidents, together with the number of reported incidents during the two-year study period. Of a total of 473 reported occupational blood exposures, the majority came from nurses and the minority from physicians. Most reported incidents occurred on hospital wards. The most common incidents were needlestick injuries, and 35% occurred when the needle was recapped. Medical laboratory technicians (MLT) reported significantly more mucocutaneous incidents than other professionals (P < 0·01). In 10% of the incidents, the patient had a known blood-borne infection. Serological investigations post-exposure varied among professional groups, and 35% were not tested. No seroconversion was shown in the HCWs tested. In the third part of the study, respondents recalled 1180 incidents, although only 9% of these had been reported. The majority occurred in operating theatres, and in connection with anaesthesia. There was a significant difference (P < 0·001) between the different professional groups with regard to the frequency of incident reporting. Physicians reported only 3% and MLTs 36% of the incidents. Eighty-one percent believed that the accident could have been avoided. Despite knowledge of universal precautions, professionals continue to behave in a risky manner, which can result in blood exposure incidents.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-84861 (URN)10.1016/S0195-6701(97)90210-3 (DOI)
    Tillgänglig från: 2012-10-25 Skapad: 2012-10-25 Senast uppdaterad: 2017-12-07Bibliografiskt granskad
    2. Health care workers' action strategies in situations that involve a risk of blood exposure
    Öppna denna publikation i ny flik eller fönster >>Health care workers' action strategies in situations that involve a risk of blood exposure
    2003 (Engelska)Ingår i: Journal of Clinical Nursing, ISSN 0962-1067, E-ISSN 1365-2702, Vol. 12, nr 5, s. 660-667Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    • Previous research shows that health care workers (HCWs) often act in a risky way in blood-exposure situations, and thereby run the risk of becoming infected by blood-borne pathogens.

    • A qualitative study was conducted in order to describe factors that influence HCWs' actions in such situations. Nurses and nursing assistants were interviewed.

    •  The analysis shows that HCWs perceive that there is a conflict among different demands. These demands are protecting the patient's privacy, protecting themselves from being infected and respecting the norms of the department.

    • The process of managing this conflict is labelled `balancing', which most often results in the choice of a diagnosis-related strategy.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26499 (URN)10.1046/j.1365-2702.2003.00644.x (DOI)11055 (Lokalt ID)11055 (Arkivnummer)11055 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    3. Blood exposure: factors promoting health care workers’ compliance with guidelines in connection with risk
    Öppna denna publikation i ny flik eller fönster >>Blood exposure: factors promoting health care workers’ compliance with guidelines in connection with risk
    2004 (Engelska)Ingår i: Journal of Clinical Nursing, ISSN 0962-1067, E-ISSN 1365-2702, Vol. 13, nr 5, s. 547-554Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Background.  Health care workers compliance with guidelines, universal precautions, in connection with tasks that could involve contact with patient's blood is unsatisfactory. In a previous paper, we identified different forces that undermine compliance. Socialization into infection control, routinization, stereotyping, perceptions of patients’ wishes and the presence of competing values and norms are examples of such forces.

    Aims and objectives.  The aim of this article is to describe and analyse different forces that promote adherence to universal precautions. Behavioural variations are seen as a consequence of differences between wards with regard to the safety culture. Safety culture is conceptualized as the outcome of a constant interplay between deactivating and reactivating forces. In this article the focus is on the latter.

    Method.  The grounded theory approach. Data were collected through interviews with nurses and assistant nurses.

    Results.  The charge nurse, informal leaders, students, infection control nurses, type of work, availability of equipment, blood-exposure incidents and media-coverage of infectious diseases are described as potentially important for compliance. The properties these agents must possess in order to be influential are also described.

    Relevance to clinical practice.  The outcome of an occupationally acquired infection can be fatal. Hence it is important that health care workers take protective measures. The results imply that mere information about safe practices alone is insufficient to achieve that goal. All factors of importance for compliance must be taken in to consideration in clinical work and in education.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-22595 (URN)10.1111/j.1365-2702.2004.00897.x (DOI)1871 (Lokalt ID)1871 (Arkivnummer)1871 (OAI)
    Tillgänglig från: 2009-10-07 Skapad: 2009-10-07 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    4. Patients’ conceptions of quality care and barrier care
    Öppna denna publikation i ny flik eller fönster >>Patients’ conceptions of quality care and barrier care
    2006 (Engelska)Ingår i: Journal of Evaluation In Clinical Practice, ISSN 1356-1294, E-ISSN 1365-2753, Vol. 12, nr 6, s. 682-691Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Aim  To describe patients’ conceptions of quality care and barrier care.

    Methods  As this study concerned conceptions of care, a phenomenographic approach was chosen. Fourteen adult orthopaedic patients participated. Data-collection was performed by means of semi-structured interviews. The qualitative data were analysed with two foci, conceptions of quality care and conceptions of barrier care. Different categories of understanding, that is, conceptions, constitute the the essential outcome of phenomenographic analysis. The research was conducted in one county hospital and in one regional hospital situated in different cities in the south of Sweden.

    Results and Conclusions  Patients’ conceptions of quality care resulted in six categories. When comparing the findings with previous research in this field, the findings of the present study confirmed to a large extent the findings from other studies of quality care. Patients’ conceptions of barrier care resulted in five categoris. The conceptions of barrier care must be considered as elements in patients’ conceptions of quality care, and this must be called atention to in efforts to measure patient satisfaction and in analyses of good care. It also can influence health care workers’ compliance to guidelines in infection control procedures.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-35740 (URN)10.1111/j.1365-2753.2006.00636.x (DOI)28386 (Lokalt ID)28386 (Arkivnummer)28386 (OAI)
    Tillgänglig från: 2009-10-10 Skapad: 2009-10-10 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
  • 58.
    Lymer, Ulla-Britt
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Antonsson Schütz, A.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Isaksson, Barbro
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    A descriptive study of blood exposure incidents among healthcare workers in a university hospital in Sweden1997Ingår i: Journal of Hospital Infection, ISSN 0195-6701, E-ISSN 1532-2939, Vol. 35, nr 3, s. 223-235Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    In an attempt to document blood exposure incidents and compliance with recommended serological investigations, universal precautions and incident reporting routines, data was collected from occupational injury reports during a two-year period. In addition, a sample of healthcare workers (HCWs) answered a questionnaire about blood tests and work routines. In a third part of the study some HCWs were asked about the type and actual frequency of incidents, together with the number of reported incidents during the two-year study period. Of a total of 473 reported occupational blood exposures, the majority came from nurses and the minority from physicians. Most reported incidents occurred on hospital wards. The most common incidents were needlestick injuries, and 35% occurred when the needle was recapped. Medical laboratory technicians (MLT) reported significantly more mucocutaneous incidents than other professionals (P < 0·01). In 10% of the incidents, the patient had a known blood-borne infection. Serological investigations post-exposure varied among professional groups, and 35% were not tested. No seroconversion was shown in the HCWs tested. In the third part of the study, respondents recalled 1180 incidents, although only 9% of these had been reported. The majority occurred in operating theatres, and in connection with anaesthesia. There was a significant difference (P < 0·001) between the different professional groups with regard to the frequency of incident reporting. Physicians reported only 3% and MLTs 36% of the incidents. Eighty-one percent believed that the accident could have been avoided. Despite knowledge of universal precautions, professionals continue to behave in a risky manner, which can result in blood exposure incidents.

  • 59.
    Lymer, Ulla-Britt
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Richt, Bengt
    Linköpings universitet, Institutionen för hälsa och samhälle. Linköpings universitet, Hälsouniversitetet.
    Patients’ conceptions of quality care and barrier care2006Ingår i: Journal of Evaluation In Clinical Practice, ISSN 1356-1294, E-ISSN 1365-2753, Vol. 12, nr 6, s. 682-691Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Aim  To describe patients’ conceptions of quality care and barrier care.

    Methods  As this study concerned conceptions of care, a phenomenographic approach was chosen. Fourteen adult orthopaedic patients participated. Data-collection was performed by means of semi-structured interviews. The qualitative data were analysed with two foci, conceptions of quality care and conceptions of barrier care. Different categories of understanding, that is, conceptions, constitute the the essential outcome of phenomenographic analysis. The research was conducted in one county hospital and in one regional hospital situated in different cities in the south of Sweden.

    Results and Conclusions  Patients’ conceptions of quality care resulted in six categories. When comparing the findings with previous research in this field, the findings of the present study confirmed to a large extent the findings from other studies of quality care. Patients’ conceptions of barrier care resulted in five categoris. The conceptions of barrier care must be considered as elements in patients’ conceptions of quality care, and this must be called atention to in efforts to measure patient satisfaction and in analyses of good care. It also can influence health care workers’ compliance to guidelines in infection control procedures.

  • 60.
    Lymer, Ulla-Britt
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Richt, Bengt
    Linköpings universitet, Institutionen för hälsa och samhälle. Linköpings universitet, Hälsouniversitetet.
    Isaksson, Barbro
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Blood exposure: factors promoting health care workers’ compliance with guidelines in connection with risk2004Ingår i: Journal of Clinical Nursing, ISSN 0962-1067, E-ISSN 1365-2702, Vol. 13, nr 5, s. 547-554Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Background.  Health care workers compliance with guidelines, universal precautions, in connection with tasks that could involve contact with patient's blood is unsatisfactory. In a previous paper, we identified different forces that undermine compliance. Socialization into infection control, routinization, stereotyping, perceptions of patients’ wishes and the presence of competing values and norms are examples of such forces.

    Aims and objectives.  The aim of this article is to describe and analyse different forces that promote adherence to universal precautions. Behavioural variations are seen as a consequence of differences between wards with regard to the safety culture. Safety culture is conceptualized as the outcome of a constant interplay between deactivating and reactivating forces. In this article the focus is on the latter.

    Method.  The grounded theory approach. Data were collected through interviews with nurses and assistant nurses.

    Results.  The charge nurse, informal leaders, students, infection control nurses, type of work, availability of equipment, blood-exposure incidents and media-coverage of infectious diseases are described as potentially important for compliance. The properties these agents must possess in order to be influential are also described.

    Relevance to clinical practice.  The outcome of an occupationally acquired infection can be fatal. Hence it is important that health care workers take protective measures. The results imply that mere information about safe practices alone is insufficient to achieve that goal. All factors of importance for compliance must be taken in to consideration in clinical work and in education.

  • 61.
    Lymer, Ulla-Britt
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Richt, Bengt
    Linköpings universitet, Institutionen för tema. Linköpings universitet, Hälsouniversitetet.
    Isaksson, Barbro
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Health care workers' action strategies in situations that involve a risk of blood exposure2003Ingår i: Journal of Clinical Nursing, ISSN 0962-1067, E-ISSN 1365-2702, Vol. 12, nr 5, s. 660-667Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    • Previous research shows that health care workers (HCWs) often act in a risky way in blood-exposure situations, and thereby run the risk of becoming infected by blood-borne pathogens.

    • A qualitative study was conducted in order to describe factors that influence HCWs' actions in such situations. Nurses and nursing assistants were interviewed.

    •  The analysis shows that HCWs perceive that there is a conflict among different demands. These demands are protecting the patient's privacy, protecting themselves from being infected and respecting the norms of the department.

    • The process of managing this conflict is labelled `balancing', which most often results in the choice of a diagnosis-related strategy.

  • 62.
    Majeed, Meytham
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Gustafsson, Mikael
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Medicinsk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Kihlström, Erik
    Department of clinical microbiology, Örebro medical center hospital, Örebro, Sweden.
    Stendahl, Olle
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Medicinsk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Roles of Ca2+ and F-actin in intracellular aggregation of Chlamydia trachomatis in eucaryotic cells1993Ingår i: Infection and Immunity, ISSN 0019-9567, E-ISSN 1098-5522, Vol. 61, nr 4, s. 1406-1414Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The effect of intracellular free Ca2+ ([Ca2+]i) on the intracellular aggregation of Chlamydia trachomatis serovars L2 and E in McCoy and HeLa cells is investigated. Loading the cells with the Ca2+ chelator MAPT/AM (1,2-bis-5-methyl-amino-phenoxylethane-N,N-n'-tetra-acetoxymethyl acetate), thereby decreasing the [Ca2+]i from 67 to 19 nM, decreased the number of cells with a local aggregation of chlamydiae in a dose-dependent manner. Neither the attachment nor the uptake of elementary bodies (EBs) was, however, affected after depletion of Ca2+ from the cells. There was no significant difference in the level of measured [Ca2+]i between infected and uninfected cells. Reducing the [Ca2+]i also significantly inhibited chlamydial inclusion formation. Differences in the organization of the actin filament network were observed in response to [Ca2+]i depletion. In Ca(2+)-depleted cells, where few EB aggregates were formed, few local accumulations of F-actin were observed in the cytosol. These results suggest that the aggregation of EBs in eucaryotic cells requires a normal homeostasis of intracellular Ca2+. By affecting F-actin reorganization and putatively certain Ca(2+)-binding proteins, [Ca2+]i plays a vital role in the infectious process of chlamydiae.

  • 63.
    Matussek, A
    et al.
    County Hospital Ryhov, Department of Clinical Microbiology, Jönköping, Sweden.
    Strindhall, J
    yDepartment of Natural Science and Biomedicine, School of Health Sciences, Jönköping, Sweden.
    Stark, Lisa
    Linköpings universitet, Institutionen för klinisk och experimentell medicin. Linköpings universitet, Hälsouniversitetet.
    Rohde, M
    zDepartment of Microbiology, German Research Centre for Biotechnology, Gemany.
    Geffers, R
    Mucosal Immunity Group, German Research Centre for Biotechnology, Braunschweig, Germany.
    Buer, J
    Mucosal Immunity Group, German Research Centre for Biotechnology, Braunschweig/Institute of Medical Microbiology, Hannover Medical School, Hannover, Germany.
    Kihlström, Erik
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Lindgren, Per-Eric
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Medicinsk mikrobiologi.
    Löfgren, S
    Department of Clinical Microbiology, County Hospital Ryhov, Jönköping.
    Infection of human endothelial cells with Staphylococcus aureus induces transcription of genes encoding an innate immunity response2005Ingår i: Scandinavian Journal of Immunology, ISSN 0300-9475, E-ISSN 1365-3083, Vol. 61, nr 6, s. 536-544Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Staphylococcus aureus is a gram-positive bacterium frequently isolated from patients with bloodstream infections. Endothelial cells (EC) play an important role in host defence against bacteria, and recent reports have shown that infection of EC with S. aureus induces expression of cytokines and cell surface receptors involved in activating the innate immune response. The ability of S. aureus to invade nonphagocytic cells, including EC, has been documented. However, the knowledge of the role of EC in pathogenesis of S. aureus infection is still limited. In this study, we investigate the gene-expression program in human EC initiated by internalized 5. aureus, using microarray analysis. We found 156 genes that were differentially regulated at least threefold, using arrays representing 14,239 genes. Many of the up regulated genes code for proteins involved in innate immunity, such as cytokines, chemokines and cell adhesion proteins. Other upregulated genes encode proteins involved in antigen presentation, cell signalling and metabolism. Furthermore, intracellular bacteria survived for days without inducing EC death. © 2005 Blackwell Publishing Ltd.

  • 64.
    Monstein, Hans-Jurg
    et al.
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för biomedicin och kirurgi, Avdelningen för medicinsk cellbiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Molekylärbiologiska tekniklaboratoriet.
    Balkhed Östholm, Åse
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Hjärt- och Medicincentrum, Infektionskliniken i Östergötland.
    Nilsson, MV
    Nilsson, M
    Dornbusch, Kathrine
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Nilsson, Lennart
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Multiplex PCR amplification assay for the detection of blaSHV, blaTEM and blaCTX-M genes in Enterobacteriaceae2007Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 115, nr 12, s. 1400-1408Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Extended-spectrum β-lactamases (ESBLs) are often mediated by bla-SHV, blaTEM and blaCTX-M genes in Enterobacteriaceae and other Gram-negative bacteria. Numerous molecular typing methods, including PCR-based assays, have been developed for their identification. To reduce the number of PCR amplifications needed we have developed a multiplex PCR assay which detects and discriminates between bla-SHV, blaTEM and blaCTX-M PCR amplicons of 747, 445 and 593 bp, respectively. This multiplex PCR assay allowed the identification of bla-SHV, blaTEM and blaCTX-M genes in a series of clinical isolates of Enterobacteriaceae with previously characterised ESBL phenotype. The presence of blaSHV, blaTEM and blaCTX-M genes was confirmed by partial DNA sequence analysis. Apparently, the universal well-established CTX-M primer pair used here to reveal plasmid-encoded blaCTX-M genes would also amplify the chromosomally located K-1 enzyme gene in all Klebsiella oxytoca strains included in the study.

  • 65.
    Monstein, Hans-Jurg
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Quenadu, Michael
    Laboratory of Food hygiene, Department of Food Thecnology, University of Lund.
    Samuelsson, Annika
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Ahrne, Siv
    Laboratory of Food hygiene, Department of Food Thecnology, University of Lund.
    Isaksson, Barbro
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Jonasson, Jon
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Division of the genus Enterococcus into species groups using PCR-based molecular typing methods1998Ingår i: Microbiology, ISSN 1350-0872, E-ISSN 1465-2080, Vol. 144, nr 5, s. 1171-1179Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Broad-range 16S rDNA PCR (BR-PCR) applied to DNA from 32 clinical enterococcal isolates and 12 other enterococci from a clinical reference collection followed by species-specific hybridization analysis identified 25 strains of Enterococcus faecalis and 19 Enterococcus species. Randomly amplified polymorphic DNA (RAPD) analysis using UPGMA clustering on the same material revealed four different clusters at a similarity level of 49%. Based on partial 16S rDNA sequence analysis of variable regions V4 and V9, it was possible to divide the 19 type strains specifying the genus Enterococcus into 12 different 16S rDNA species groups. The type strain distribution then served as a template for the analysis of the other 44 strains which were assigned to four different species groups (a-d) based on their 16S rDNA motifs. There was good agreement with the RAPD clusters. Species group a was an individual species line containing 25 strains that were identified as E. faecalis. Group b also represented an individual species line of 12 strains identified as E. faecium. The remaining seven strains that formed species groups c and d could not be fully identified to species by this analysis. It was concluded that BR-PCR of 16S rDNA followed by partial sequence analysis of the PCR products is a reliable technique for the identification and classification of enterococci. Further division of unresolved species groups should be achievable if regions other than V4 and V9 of 16S rDNA are also analysed

  • 66.
    Monstein, Hans-Jurg
    et al.
    Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Tiveljung, Annika
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Kraft, C. H.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Borch, Kurt
    Linköpings universitet, Institutionen för biomedicin och kirurgi, Kirurgi. Linköpings universitet, Hälsouniversitetet.
    Jonasson, Jon
    Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Profiling of bacterial flora in gastric biopsies from patients with Helicobacter pylori-associated gastritis and histologically normal control individuals by temperature gradient gel electrophoresis and 16S rDNA sequence analysis2000Ingår i: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 49, nr 9, s. 817-822Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The aim of this study was to establish bacterial profiles in gastric biopsy specimens from patients with Helicobacter pylori-associated gastritis by means of temporal temperature gradient gel electrophoresis (TTGE) of PCR-amplified 16S rDNA fragments. Specimens from eight patients with asymptomatic gastritis and five histologically normal controls revealed a Helicobacter-specific band in the TTGE profile with increased amounts of Helicobacter-specific DNA in the biopsies from most of the gastritis patients. DNA from other genera including Enterococcus, Pseudomonas, Streptococcus, Staphylococcus and Stomatococcus was also found in the stomach. In the absence of gastric inflammation, Helicobacter spp. appeared to be part of a complex, presumably indigenous microbial flora found in the biopsy specimens from the stomach.

  • 67.
    Monstein, Hans-Jürg
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet. Östergötlands Läns Landsting, Diagnostikcentrum, Klinisk mikrobiologi.
    Kihlström, Erik
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet. Östergötlands Läns Landsting, Diagnostikcentrum, Klinisk mikrobiologi.
    Tiveljung, Annika
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Detection and identification of bacteria using in-house broad range 16S rDNA PCR amplification and genus-specific DNA hybridization probes, located within variable regions of 16S rRNA genes1996Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 104, nr 1-6, s. 451-458Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Broad range PCR amplification and genus-specific 16S ribosomal DNA hybridization was used to demonstrate that Chlamydia, Helicobacter and Mobiluncus hybridization probes, located within variable regions V3, V4, and V9 of the 16S rDNA, specifically bound to the corresponding PCR product obtained from pure cultures of the three genera. The sensitivity of the assay was determined by analysis of C. trachomatis serially diluted in urine. The detection limit was 1–10 elementary bodies using a hybridization probe derived from the variable region V3 of the 16S rRNA gene. A PCR product was furthermore formed in urine specimens not containing C. trachomatis, showing amplification of chlamydia also in the presence of DNA from the resident urethral flora that competes for annealing sites. Analysis of a restricted number of male urine specimens using the C. trachomatis-specific probe showed complete agreement with culture and a commercially available PCR kit. Our method not only has the capacity to detect C. trachomatis in microbiologically mixed urine samples but also the potential advantage of identifying other bacterial pathogens from the same PCR product by varying the hybridization probes.

  • 68.
    Monstein, H.-J.
    et al.
    Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Nilsson, Isabelle
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Ellnebo-Svedlund, Katarina
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Svensson, S.P.S.
    Linköpings universitet, Institutionen för medicin och vård, Farmakologi. Linköpings universitet, Hälsouniversitetet.
    Cloning and characterization of 5'-end alternatively spliced human cholecystokinin-B receptor mRNAs1998Ingår i: Receptors and Channels, ISSN 1060-6823, E-ISSN 1607-856X, Vol. 6, nr 3, s. 165-177Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    We report here the cloning and characterization of a 5'-end alternatively spliced human cholecystokinin-B (CCK-B) receptor mRNA. The 5'-end of this CCK-B receptor transcript (termed CCK-BRtx) consisted of exon Ia, present in the ordinary full-length CCK-B receptor mRNA (CCK-BRwt), and exon Ib, present in a previously described 5'-end alternatively spliced CCK-B receptor mRNA (CCK-BRt). A short open reading frame preceded the AUG translation initiation codon of the CCK-BRtx. Transfection of COS-7 cells with the CCK-BRtx or CCK-BRt cDNAs did not lead to the appearance of peptidergic and non-peptidergic binding sites. Cell free in vitro translation yielded proteins of approximately 44 kDa (CCK-B receptor) and 40 kDa (CCK-BRt receptor) whereas no 40 kDa product was detected from the cloned CCK-BRtx cDNA. Instead, a protein product of approximately 9 kDa was visualized, the size corresponding to the predicted protein encoded by the short open reading frame. The alternatively spliced CCK-B receptor transcripts were concomitantly expressed with the ordinary full-length CCK-B receptor mRNA in the brain, pancreas, and stomach. The possibility that such transcripts are translated in vivo into truncated CCK-B receptors is discussed.

  • 69.
    Mölling, Paula
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Genetic characterisation of Meningococci2001Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [sv]

    Meningokockmeningit och/eller sepsis är infektion på hjärnhinnan (hjärnhinneinflammation) och/eller i blodet orsakad av Neisseria meningitidis bakterier, även kallade meningekocker (Mc). Sjukdomen kan framkalla stor rädsla hos många föräldrar, familjemedlemmar och av vårdpersonal eftersom den ofta utvecklas snabbt, har en hög dödlighet och kan vara svår att skilja från andra febersjukdomar. De som överlever kan ha fått permanenta vävnadsskador och neurologiska problem. Den snabba utvecklingen av sjukdomen hos vanligen friska barn eller ungdomar har resulterat i intensiv forskning inom området. Speciellt gäller det diagnostik och karakteriseringsmetoder, vaccinutveckling, samt vad det är som gör vissa sorter av Mc mer virulenta (sjukdomsframkallande) än andra. Denna avhandling (Papper I-VI) handlar om utvecklingen av genetiska metoder för att både mer fullständigt och snabbare kunna identifiera och karakterisera Mc.

    Mc sjukdom diagnostiseras vanligen genom att man odlar fram bakterien, men p.g.a. av det ökande användandet av antibiotika och ev. transportproblem kan många prov bli falskt "negativa". För att identifiera de mest sjukdomsframkallande grupperna (A, B, C, Y och W-135) har genetiska metoder nu utvecklats/vidareutvecklats (Papper I, II). Bakterien delas även in i subtyper, vilka också inverkar på bakteriens virulens. Vanligen görs denna karakterisering från odlade bakterier men man har mer och mer övergått till att utgå från bakteriens DNA (genosubtypning) för att få en fullständig karakterisering (Papper III-V). För epidemiologiska syften kan genesubtypning användas tillsammans med en metod där bakteriens DNA delas upp i småbitar för att få ett s.k. "fingeravtryck" av bakterien (Papper IV-V).

    I papper VI presenteras den första DNA sekvensen på en ovanligt förekommande Mc plasmid, innehållande genen för ß-laktamas, vilket gör det möjligt för bakterien att byta ner penicillin-antibiotika och gör den därmed resistent. Plasmiden visade sig vara en variant av en plasmid man hittat hos gonokocker (Ge), en nära släkting till Mc, vilket gör det troligt att plasmiden har överförts från Ge. Blir dessa plasmider vanligare hos Mc skulle en förändring av nuvarande antibiotikabehandling av Mc sjukdom behöva göras.

    Delarbeten
    1. PCR Identification of the Group A Neisseria Meningitidis Gene in Cerebrospinal Fluid
    Öppna denna publikation i ny flik eller fönster >>PCR Identification of the Group A Neisseria Meningitidis Gene in Cerebrospinal Fluid
    1999 (Engelska)Ingår i: Scandinavian Journal of Infectious Diseases, ISSN 0036-5548, E-ISSN 1651-1980, Vol. 31, nr 5, s. 481-483Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The aim of this study was to develop a PCR method for direct identification of Neisseria meningitidis serogroup A in cerebrospinal fluid. The assay makes use of unique sites within the gene cassette responsible for expression of the (α1→6)-linked N-acetyl-D-mannosamine-1-phosphate serogroup A capsule. A total of 67 different N. meningitidis strains and 12 clinical samples of CSF, culture positive for N. meningitidis, were examined. All the strains and samples of N. meningitidis serogroup A were correctly identified by an amplified PCR product of 519 bp. The PCR method for identification is specific for the group A gene of N. meningitidis. The assay may contribute to reducing recurrent, devastating epidemics of meningococcal infection by providing a diagnostic tool for grouping in developing countries where problems with false negative cultures are common and vaccination against serogroup A meningococci may be required.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26071 (URN)10.1080/00365549950164003 (DOI)10530 (Lokalt ID)10530 (Arkivnummer)10530 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    2. Direct and Rapid Identification and Genogrouping of Meningococci and porA Amplification by LightCycler PCR
    Öppna denna publikation i ny flik eller fönster >>Direct and Rapid Identification and Genogrouping of Meningococci and porA Amplification by LightCycler PCR
    2002 (Engelska)Ingår i: Journal of Clinical Microbiology, ISSN 0095-1137, E-ISSN 1098-660X, Vol. 40, nr 12, s. 4531-4535Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Invasive meningococcal infections are usually diagnosed by culture. This approach is far from optimal due to, e.g., treatment with precollection antibiotics. Molecular-genetics methods are therefore recognized as important tools for optimal laboratory confirmation of meningococcal infections as well as characterization of meningococci (Mc). The aims of the present study were to develop real-time PCRs for identification and genogrouping (A, B, C, Y, and W-135) of Mc and porA amplification that further can be used for subsequent genosubtyping. In a first run Mc were identified. In a second run they were genogrouped and porA genes were amplified. All the Mc isolates (n = 71) but one and cerebrospinal fluid samples (n = 11) tested gave the expected positive results. The specificity, inter- and intra-assay variations, and effects of different amounts of DNA on the melting temperatures were also explored. The LightCycler PCR system was found to effectively detect and characterize Mc in a few hours. This testing, including the DNA sequencing of the porA gene to reveal the genosubtype, does not take more than a working day, and the results can be compared to those from culturing with phenotypic analysis, which takes at least a few days.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26553 (URN)10.1128/​JCM.40.12.4531-4535.2002 (DOI)11115 (Lokalt ID)11115 (Arkivnummer)11115 (OAI)
    Anmärkning
    On the day of the defence day the status of this article was a manuscript.Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    3. Genosubtyping by sequencing group A, B and C meningococci: a tool for epidemiological studies of epidemics, clusters and sporadic cases
    Öppna denna publikation i ny flik eller fönster >>Genosubtyping by sequencing group A, B and C meningococci: a tool for epidemiological studies of epidemics, clusters and sporadic cases
    2000 (Engelska)Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 108, nr 7-8, s. 509-516Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Genosubtyping, by sequencing variable regions (VRs) 1, 2 and 3 of the porA gene, was evaluated as a tool to detect clonality of isolates in meningococcal epidemics in Africa and clusters of disease in Sweden. All 63 examined meningococcal isolates were successfully genosubtyped. The isolates belonging to group A type 4 with genosubtype P1.20,9,35a showed little heterogeneity in African epidemics in 1988 and onwards. In Sweden, two meningococcal clones of group B type 15, with genosubtypes P1.7,16,35 and P1.7,16f,35, dominated during two clusters of meningococcal disease in 1995–96 and in sporadic cases thereafter. The characterisation of group C meningococci isolated during 1992 in Sweden indicated a cluster (type 2a with genosubtype P1.5a,10d,36b) connected with a discotheque visit. Two variants of VR2 (10p and 25b), not previously described, were found among the examined isolates. Nucleotide sequence analysis of VRs in the porA gene proved a valuable epidemiological tool since almost all isolates could be genosubtyped, in contrast to the phenotypic methods presently used.

    Nyckelord
    Epidemiology, genosubtyping, sequencing, Neisseria meningitidis, meningococcal disease
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-25867 (URN)10.1034/j.1600-0463.2000.01087-8509.x (DOI)10304 (Lokalt ID)10304 (Arkivnummer)10304 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    4. Comparison of Serogroup W-135 Meningococci Isolated in Sweden during a 23-Year Period and Those Associated with a Recent Hajj Pilgrimag
    Öppna denna publikation i ny flik eller fönster >>Comparison of Serogroup W-135 Meningococci Isolated in Sweden during a 23-Year Period and Those Associated with a Recent Hajj Pilgrimag
    2001 (Engelska)Ingår i: Journal of Clinical Microbiology, ISSN 0095-1137, E-ISSN 1098-660X, Vol. 39, nr 7, s. 2695-2699Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    An outbreak of serogroup W-135 meningococcal disease was reported among pilgrims returning from the annual hajj (pilgrimage to Mecca) in mid-March 2000. Molecular characterization was used to investigate the similarity of the hajj-associated W-135 strains with those isolated in Sweden during a 23-year period (1978 to 2000). The same hajj-associated genosubtype, genosubtype P1.5,2,36b, has been documented in Sweden since 1979, while pulsed-field gel electrophoresis and the sulfadiazine resistance of the W-135 isolates indicated that the outbreak was probably due to a new clone of W-135 meningococci.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-25865 (URN)10.1128/JCM.39.7.2695-2699.2001 (DOI)10302 (Lokalt ID)10302 (Arkivnummer)10302 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    5. Long-Term Persistence of a Discotheque-Associated Invasive Neisseria meningitidis Group C Strain as Proven by Pulsed-Field Gel Electrophoresis and porA Gene Sequencin
    Öppna denna publikation i ny flik eller fönster >>Long-Term Persistence of a Discotheque-Associated Invasive Neisseria meningitidis Group C Strain as Proven by Pulsed-Field Gel Electrophoresis and porA Gene Sequencin
    2000 (Engelska)Ingår i: Journal of Clinical Microbiology, ISSN 0095-1137, E-ISSN 1098-660X, Vol. 38, nr 4, s. 1638-1640Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    A cluster of a Neisseria meningitidis serogroup C strain causing invasive disease was investigated. Five out of seven cases were associated with a particular discotheque. The strains were indistinguishable, as revealed by pulsed-field gel electrophoresis and sequencing of variable regions of the porA gene, but caused strikingly different clinical presentations during 5 months.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-25919 (URN)10361 (Lokalt ID)10361 (Arkivnummer)10361 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    6. Complete Sequence of a β-Lactamase-Encoding Plasmid in Neisseria meningitidis
    Öppna denna publikation i ny flik eller fönster >>Complete Sequence of a β-Lactamase-Encoding Plasmid in Neisseria meningitidis
    Visa övriga...
    2000 (Engelska)Ingår i: Antimicrobial Agents and Chemotherapy, ISSN 0066-4804, E-ISSN 1098-6596, Vol. 44, nr 1, s. 210-212Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Identical β-lactamase-encoding (TEM-1) plasmids were found in two different clinical Neisseria meningitidis strains. They were completely sequenced (5,597 bp) and designated pAB6. The plasmid is almost identical to Neisseria gonorrhoeae plasmid pJD5 (5,599 kb) and may have been picked up from a gonococcus in vivo.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-25866 (URN)10.1128/​AAC.44.1.210-212.2000 (DOI)10303 (Lokalt ID)10303 (Arkivnummer)10303 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
  • 70.
    Nayeri, Fariba
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    Aili, Daniel
    Linköpings universitet, Institutionen för fysik, kemi och biologi. Linköpings universitet, Tekniska högskolan.
    Nayeri, Tayeb
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    Xu, Junyang
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    Almer, Sven
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Gastroenterologi och hepatologi. Linköpings universitet, Hälsouniversitetet.
    Lundström, Ingemar
    Linköpings universitet, Institutionen för fysik, kemi och biologi. Linköpings universitet, Tekniska högskolan.
    Åkerlind, Britt
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Liedberg, Bo
    Linköpings universitet, Institutionen för fysik, kemi och biologi. Linköpings universitet, Hälsouniversitetet.
    Hepatocyte growth factor (HGF) in fecal samples: rapid detection by surface plasmon resonance2005Ingår i: BMC Gastroenterology, ISSN 1471-230X, E-ISSN 1471-230X, Vol. 5, nr 13Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Background

    The development of biosensors, based on surface plasmon resonance (SPR) technology, enables monitoring of a variety of biospecific interactions without the need for chemical-, biological- or radiological-labelled reagents.

    Method

    We utilised SPR to detect hepatocyte growth factor (HGF) in reconstituted faecal samples and studied samples from patients with infectious gastroenteritis (n = 20) and normal controls (n = 10). Mouse anti-human HGF monoclonal antibodies and recombinant human HGF receptor (c-Met)/Fc chimera were immobilised in flow cells of a CM5 biosensor chip.

    Results

    We found that infectious gastroenteritis produced a higher signal response compared to controls, due to binding of HGF to monoclonal anti-HGF antibody as well as binding of HGF to c-Met receptor (p < 0.01). The SPR signal response correlated with results from ELISA (r = 72%, p > 0.001). The signal response decreased significantly (p < 0.05) when samples were diluted with dextran, because of reduction in both specific as well as unspecific binding of HGF to dextran. The decrease in the specific response might imply that the dextran- binding site for HGF overlaps with the antibody binding epitope, or that dextran binding induces a conformational change of the HGF molecule. Bands corresponding to HGF were found by gel electrophoresis of purified faeces in an affinity chromatography column immobilised by HGF ligands.

    Conclusion

    Determination of HGF by SPR might be beneficial in diagnosis of acute situations that present with symptoms of gastroenteritis and may, possibly, guide appropriate medical treatments. This is to our knowledge the first report on the use of SPR for detection of HGF in faeces samples.

  • 71.
    Nayeri, Fariba
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    Almer, Sven
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Gastroenterologi och hepatologi. Linköpings universitet, Hälsouniversitetet.
    Brudin, Lars
    Department of Clinical Physiology, County Hospital, Kalmar.
    Nilsson, Ingela
    Department of Clinical Chemistry, County Hospital, Kalmar.
    Åkerlind, Britt
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Forsberg, Pia
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    High hepatocyte growth factor levels in faeces during acute infectious gastroenteritis2003Ingår i: Scandinavian Journal of Infectious Diseases, ISSN 0036-5548, E-ISSN 1651-1980, Vol. 35, nr 11-12, s. 858-862Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Hepatocyte growth factor (HGF) is a potent mitogen of mature epithelial cells which is produced after organ injuries and acts as a trigger for regeneration in the impaired organ. The aim of the present study was to investigate local production of HGF during infectious gastroenteritis. We measured the concentration of HGF in serum and faeces in 49 patients with acute infectious gastroenteritis (bacterium=30, virus=10, amoebae=1, and probable infection=8) at the time of referral to hospital and at convalescence (n=31). The values were compared with normal healthy vaccination volunteers (n=11) as well as patients with acute non-infectious diarrhoea (n=10). The presence of HGF in faeces was confirmed by ELISA and Western immunoblot. HGF concentrations in faeces was significantly higher in the patients with infectious gastroenteritis compared to the control groups (p<0.0001). Using a cut-off concentration of 20 ng/g, the overall sensitivity of faeces HGF to distinguish infectious gastroenteritis (bacterial, viral, probable infection) was 98% with a specificity of 100%. At convalescence all patients had normal values. There was no significant correlation between HGF concentrations in faeces and serum. Determination of faeces HGF may identify cases of transmittable diarrhoea requiring isolation at an early stage.

  • 72.
    Nordin, Gunnar
    et al.
    Uppsala .
    Klinteberg, Barbro
    Helsingborg .
    Persson, Birgitta
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Forsum, Urban
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Får en laboratorieundersökning kallas vad som helst?2005Ingår i: Läkartidningen, ISSN 0023-7205, E-ISSN 1652-7518, Vol. 102, nr 17, s. 1308-1315Artikel i tidskrift (Övrigt vetenskapligt)
  • 73.
    Richt, Bengt
    et al.
    Linköpings universitet, Filosofiska fakulteten. Linköpings universitet, Institutionen för hälsa och samhälle, Tema hälsa och samhälle.
    Lymer, Ulla-Britt
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Mötet med den farliga kroppen2004Ingår i: När människan möter medicinen.: Livsvärldens och berättelsens betydelse för förståelsen av sjukdom och medicinsk teknologi. / [ed] Sonja Olin Lauritzen, Fredrik Svenaeus & Ann-Christine Jonsson, Stockholm: Carlssons förlag , 2004, 1, s. 163-188Kapitel i bok, del av antologi (Övrigt vetenskapligt)
    Abstract [sv]

    När möter människan medicinen? Det sker varje gång en individ söker sig till hälso- och sjukvården för att få hjälp med sina besvär. Denna antologi fokuserar på kommunikationer mellan människor och mellan olika kunskapstraditioner i vården, patientens förståelse av diagnosen, och på den nya medicinska teknologins konsekvenser för människor i deras dagliga liv .

  • 74.
    Rytkönen, Anne
    et al.
    KI.
    Albinger, Barbara
    Hansson-Palo, Paola
    Källström, Helena
    Olcén, Per
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Fredlund, Hans
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Jonsson, Ann-Beth
    Neisseria meningitidis undergoes PilC phase variation and PilC sequence variation during invasive disease.2004Ingår i: Journal of Infectious Diseases, ISSN 0022-1899, E-ISSN 1537-6613, Vol. 189, s. 402-409Artikel i tidskrift (Refereegranskat)
  • 75.
    Saeedi, Baharak
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    Characterization of clinical enterococcal isolates in Swedish hospitals: studies on genetic relatedness and high-level gentamicin resistance2005Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    During the last few decades, and in parallel with increasing resistance to multiple antibiotics, enterococci have become one of the leading pathogens that cause nosocomial infections. High-level gentamicin resistant (HLGR) enterococci have become frequent. Thus, there are compelling reasons to control the transmission of enterococci with HLGR. Many different typing methods have been used for epidemiological typing of enterococci. Pulsed-field gel electrophoresis (PFGE) has been shown to be the most discriminating typing method and is currently considered the "gold standard" for typing of enterococci. However, PFGE is an expensive method and remains time-consuming, which may be of critical importance when comparing data obtained from numerous isolates.

    The aim of this thesis was to characterize clinical enterococcal isolates from patients admitted to Swedish hospitals, with special focus on HLGR strains and their genetic relatedness. Our purpose was also to develop a faster PFGE protocol for typing of enterococci, as well as to investigate if the Phene Plate (PhP) system can be used as a rapid screening method for detection of genetically related isolates of enterococci. If this could be achieved, it would be possible to minimize the number of isolates subjected to PFGE typing, which would save time and money.

    In paper I, we performed a thorough investigation of eight different parameters of importance for the separation of DNA fragments by PFGE. This resulted in a modified PFGE protocol for typing of enterococci with much enhanced resolution. HLGR E. faecalis isolates obtained from patients admitted to eight Swedish ICUs during 1996 and 1998 (paper II), and isolates obtained from patients with bacteremia in the County of Östergötland during the period 1994-2001 (paper III) were characterized by our modified PFGE method. We found that the majority (69%) of isolates from ICUs in the eastern and central parts of southern Sweden, belonged to one dominating cluster, and the same cluster was also found in blood isolates from Östergötland. In nearly all cases, HLGR was due to the presence of the aac(6 ')Ie-aph(2 '')la gene situated on a Tn5281-like transposon (paper II). In the County of Östergötland, HLGR E. faecalis was first isolated from blood cultures in 1996, and the first blood isolates of HLGR E. faecium were found in 1999. During the study period, only 4 HLGR E. faecium isolates were observed, and all of them showed unique PFGE patterns.

    To evaluate the efficiency of the gentamicin disk diffusion method for detection of HLGR in clinical isolates of enterococci, all enterococcal blood isolates from paper III were studied with a 30-µg gentamicin disk as recommended by SRGA, and the method was found to have 100% sensitivity and specificity when compared with PCR.

    A "biochemical fingerprinting" method (PhP) was compared with PFGE for epidemiological characterization of enterococci. In earlier studies of the PhP method, enterococci were collected mainly from the environment or from normal human flora. Our study indicates that PhP may be a useful screening method for clinical E. faecalis isolates, with a relatively high concordance with PFGE, but that it is less useful for E. faecium since the concordance for E. faecium was low. To fully evaluate PhP as a tool for epidemiological characterization of enterococci from clinical settings, and to address questions regarding the validity of PFGE, further studies using additional genotyping methods, preferably newer typing systems based on DNA sequencing such as MLST, are warranted.

    Delarbeten
    1. Modified pulsed-field gel electrophoresis protocol for typing of enterococci
    Öppna denna publikation i ny flik eller fönster >>Modified pulsed-field gel electrophoresis protocol for typing of enterococci
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    2002 (Engelska)Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 110, nr 12, s. 869-874Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Controlling the spread of vancomycin-resistant enterococci (VRE) is an important task in hospital epidemiology. Pulsed-field gel electrophoresis (PFGE) has become the golden standard for molecular epidemiological characterisation of enterococcal isolates. For separation of DNA fragments by PFGE, different electrophoresis conditions have been recommended, but none of these protocols allows a satisfactory separation of both small and large DNA fragments of enterococci simultaneously. In this study we have speeded up the preparation of chromosomal DNA and defined new electrophoresis conditions that enhance separation of small and large DNA fragments for subtyping of enterococci with a 24 h PFGE.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26482 (URN)10.1034/j.1600-0463.2002.1101205.x (DOI)11034 (Lokalt ID)11034 (Arkivnummer)11034 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    2. Genetic relatedness among Enterococcus faecalis with transposon-mediated high-level gentamicin resistance in Swedish intensive care units
    Öppna denna publikation i ny flik eller fönster >>Genetic relatedness among Enterococcus faecalis with transposon-mediated high-level gentamicin resistance in Swedish intensive care units
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    2003 (Engelska)Ingår i: Journal of Antimicrobial Chemotherapy, ISSN 0305-7453, E-ISSN 1460-2091, Vol. 52, nr 2, s. 162-167Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    We studied 45 isolates of Enterococcus faecalis with high-level gentamicin resistance (HLGR), all but one concomitantly resistant to ciprofloxacin, and 25 ciprofloxacin-resistant isolates without HLGR for genetic relatedness using pulsed-field gel electrophoresis (PFGE). E. faecalis were isolated from patients admitted to intensive care units at eight hospitals in southern Sweden from December 1996 through December 1998. Genomic analysis by PFGE resulted in three clusters of genetically related isolates (designated clusters I, II and III) and 23 unique clones. Cluster I was found predominantly in the eastern and central parts of southern Sweden and clusters II and III in south-western Sweden. Among the 45 isolates with HLGR, 69% belonged to cluster I, 20% to cluster II, and 11% had unique PFGE patterns, which suggests that the majority of isolates with HLGR are closely related. Among the 25 ciprofloxacin-resistant isolates without HLGR, 68% had unique PFGE patterns, 12% belonged to cluster I and 20% to cluster III, which suggests the ciprofloxacin-resistant isolates are not related. All isolates with HLGR contained the aac(6)Ie-aph(2)Ia gene, which was carried on a Tn5281-like transposon in all isolates except one. We conclude that HLGR in E. faecalis was mainly due to dissemination of genetically related clones during the time studied, and that HLGR in these isolates was due to the presence of the aac(6)Ie-aph(2)Ia gene.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26483 (URN)10.1093/jac/dkg315 (DOI)11035 (Lokalt ID)11035 (Arkivnummer)11035 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    3. Genetic relatedness of Enterococcus faecalis isolates with high-level gentamicin resistance from patients with bacteraemia in the south east of Sweden 1994-2001
    Öppna denna publikation i ny flik eller fönster >>Genetic relatedness of Enterococcus faecalis isolates with high-level gentamicin resistance from patients with bacteraemia in the south east of Sweden 1994-2001
    Visa övriga...
    2004 (Engelska)Ingår i: Scandinavian Journal of Infectious Diseases, ISSN 0036-5548, E-ISSN 1651-1980, Vol. 36, nr 6-7, s. 405-409Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    High-level gentamicin resistant (HLGR) enterococci (Enterococcus faecalis and Enterococcus faecium) have become a substantial nosocomial problem in many countries. In this study, we investigated the prevalence of HLGR enterococci and their genetic relatedness in blood culture isolates from patients with bacteraemia admitted to the 3 hospitals in Östergötland, a county in the south east of Sweden, during 1994–2001. 36 of 250 E. faecalis (14%) and 4 of 106 E. faecium isolates (4%) were shown by PCR to carry the aac(6′)-Ie-aph(2″)-Ia aminoglycoside modifying gene and these isolates were also classified as HLGR enterococci by the gentamicin antibiotic disk diffusion method. A majority of HLGR E. faecalis isolates (83%) belonged to the same cluster of genetically related isolates, according to the pulsed-field gel electrophoresis (PFGE) patterns, whereas all 4 HLGR E. faecium isolates had unique PFGE patterns. In conclusion, our study showed that in contrast to studies from many other countries, the presence of HLGR enterococci was more common in E. faecalis than in E. faecium and appeared the first time in 1996 and 1999, respectively. Bacteraemia with HLGR enterococci in Östergötland was mainly due to the spread of a cluster related of E. faecalis strains.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-24621 (URN)10.1080/00365540410020622 (DOI)6802 (Lokalt ID)6802 (Arkivnummer)6802 (OAI)
    Tillgänglig från: 2009-10-07 Skapad: 2009-10-07 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    4. Phene Plate (PhP) biochemical fingerprinting: a screening method for epidemiological typing of enterococcal isolates?
    Öppna denna publikation i ny flik eller fönster >>Phene Plate (PhP) biochemical fingerprinting: a screening method for epidemiological typing of enterococcal isolates?
    Visa övriga...
    2005 (Engelska)Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 113, nr 9, s. 603-612Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Pulsed-field gel electrophoresis (PFGE) is currently considered the gold standard for genotyping of enterococci. However, PFGE is both expensive and time-consuming. The purpose of this study was to investigate whether the PhP system can be used as a reliable clinical screening method for detection of genetically related isolates of enterococci. If so, it should be possible to minimize the number of isolates subjected to PFGE typing, which would save time and money. Ninety-nine clinical enterococcal isolates were analysed by PhP (similarity levels 0.90–0.975) and PFGE (similarity levels ≤3 and ≤6 bands) and all possible pairs of isolates were cross-classified as matched or mismatched. We found that the probability that a pair of isolates (A and B) belonging to the same type according to PhP also belong to the same cluster according to PFGE, i.e. p(APFGE=BPFGE • APhP=BPhP), and the probability that a pair of isolates of different types according to PhP also belong to different clusters according to PFGE, i.e. p(APFGE≠BPFGE • APhP≠BPhP), was relatively high for E. faecalis (0.86 and 0.96, respectively), but was lower for E. faecium (0.51 and 0.77, respectively). The concordance which shows the probability that PhP and PFGE agree on match or mismatch was 86%–93% for E. faecalis and 54%–66% for E. faecium, which indicates that the PhP method may be useful for epidemiological typing of E. faecalis in the current settings but not for E. faecium.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-33468 (URN)10.1111/j.1600-0463.2005.apm_217.x (DOI)19490 (Lokalt ID)19490 (Arkivnummer)19490 (OAI)
    Tillgänglig från: 2009-10-09 Skapad: 2009-10-09 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
  • 76.
    Saeedi, Baharak
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Hällgren, Anita
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Isaksson, Barbro
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Jonasson, Jon
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Nilsson, Lennart
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Hanberger, Håkan
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    Genetic relatedness of Enterococcus faecalis isolates with high-level gentamicin resistance from patients with bacteraemia in the south east of Sweden 1994-20012004Ingår i: Scandinavian Journal of Infectious Diseases, ISSN 0036-5548, E-ISSN 1651-1980, Vol. 36, nr 6-7, s. 405-409Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    High-level gentamicin resistant (HLGR) enterococci (Enterococcus faecalis and Enterococcus faecium) have become a substantial nosocomial problem in many countries. In this study, we investigated the prevalence of HLGR enterococci and their genetic relatedness in blood culture isolates from patients with bacteraemia admitted to the 3 hospitals in Östergötland, a county in the south east of Sweden, during 1994–2001. 36 of 250 E. faecalis (14%) and 4 of 106 E. faecium isolates (4%) were shown by PCR to carry the aac(6′)-Ie-aph(2″)-Ia aminoglycoside modifying gene and these isolates were also classified as HLGR enterococci by the gentamicin antibiotic disk diffusion method. A majority of HLGR E. faecalis isolates (83%) belonged to the same cluster of genetically related isolates, according to the pulsed-field gel electrophoresis (PFGE) patterns, whereas all 4 HLGR E. faecium isolates had unique PFGE patterns. In conclusion, our study showed that in contrast to studies from many other countries, the presence of HLGR enterococci was more common in E. faecalis than in E. faecium and appeared the first time in 1996 and 1999, respectively. Bacteraemia with HLGR enterococci in Östergötland was mainly due to the spread of a cluster related of E. faecalis strains.

  • 77.
    Saeedi, Baharak
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Hällgren, Anita
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Jonasson, Jon
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Nilsson, Lennart
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Hanberger, Håkan
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    Isaksson, Barbro
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Modified pulsed-field gel electrophoresis protocol for typing of enterococci2002Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 110, nr 12, s. 869-874Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Controlling the spread of vancomycin-resistant enterococci (VRE) is an important task in hospital epidemiology. Pulsed-field gel electrophoresis (PFGE) has become the golden standard for molecular epidemiological characterisation of enterococcal isolates. For separation of DNA fragments by PFGE, different electrophoresis conditions have been recommended, but none of these protocols allows a satisfactory separation of both small and large DNA fragments of enterococci simultaneously. In this study we have speeded up the preparation of chromosomal DNA and defined new electrophoresis conditions that enhance separation of small and large DNA fragments for subtyping of enterococci with a 24 h PFGE.

  • 78.
    Saeedi, Baharak
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Tärnberg, Maria
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Gill, Hans
    Linköpings universitet, Institutionen för medicinsk teknik. Linköpings universitet, Tekniska högskolan.
    Hällgren, Anita
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Jonasson, Jon
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Nilsson, Lennart
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Isaksson, Barbro
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Kühn, I.
    Department of Microbiology and Tumour Biology Centre, Karolinska Institute, Stockholm, Sweden.
    Hanberger, Håkan
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    Phene Plate (PhP) biochemical fingerprinting: a screening method for epidemiological typing of enterococcal isolates?2005Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 113, nr 9, s. 603-612Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Pulsed-field gel electrophoresis (PFGE) is currently considered the gold standard for genotyping of enterococci. However, PFGE is both expensive and time-consuming. The purpose of this study was to investigate whether the PhP system can be used as a reliable clinical screening method for detection of genetically related isolates of enterococci. If so, it should be possible to minimize the number of isolates subjected to PFGE typing, which would save time and money. Ninety-nine clinical enterococcal isolates were analysed by PhP (similarity levels 0.90–0.975) and PFGE (similarity levels ≤3 and ≤6 bands) and all possible pairs of isolates were cross-classified as matched or mismatched. We found that the probability that a pair of isolates (A and B) belonging to the same type according to PhP also belong to the same cluster according to PFGE, i.e. p(APFGE=BPFGE • APhP=BPhP), and the probability that a pair of isolates of different types according to PhP also belong to different clusters according to PFGE, i.e. p(APFGE≠BPFGE • APhP≠BPhP), was relatively high for E. faecalis (0.86 and 0.96, respectively), but was lower for E. faecium (0.51 and 0.77, respectively). The concordance which shows the probability that PhP and PFGE agree on match or mismatch was 86%–93% for E. faecalis and 54%–66% for E. faecium, which indicates that the PhP method may be useful for epidemiological typing of E. faecalis in the current settings but not for E. faecium.

  • 79.
    Schöier, Johan
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Öllinger, Karin
    Linköpings universitet, Institutionen för nervsystem och rörelseorgan, Patologi. Linköpings universitet, Hälsouniversitetet.
    Kvarnström, Maria
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk immunologi. Linköpings universitet, Hälsouniversitetet.
    Söderlund, Gustaf
    Linköpings universitet, Institutionen för biomedicin och kirurgi, Onkologi. Linköpings universitet, Hälsouniversitetet.
    Kihlström, Erik
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Chlamydia trachomatis -induced apoptosis occurs in uninfected McCoy cells late in the developmental cycle and is regulated by the intracellular redox state2001Ingår i: Microbial Pathogenesis, ISSN 0882-4010, E-ISSN 1096-1208, Vol. 31, nr 4, s. 173-184Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Infections with the obligate intracellular bacterium Chlamydia trachomatis are characterized by avoidance of fusion between chlamydia-containing endosomes and lysosomes, bacterial persistence and development of post-infectious sequelae. In this report we show that C. trachomatis induces apoptosis in McCoy and HeLa cells. Apoptosis was monitored by three different techniques; enzyme-linked immunoassay (EIA) of fragmented nucleosomes, terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) and flow cytometry of propidium iodide-stained cells. Apoptosis occurred in uninfected cells, was induced late in the chlamydial developmental cycle, beyond 24 h post-infection and was dependent on bacterial protein synthesis. Apoptosis was not significantly increased in infected, inclusion-containing cells. Treatment of cells with the antioxidants ascorbic acid (10 μM) and α-tocopherol (10 μM) reduced the degree of apoptosis. These results suggest that host cells infected with C. trachomatis generate proapoptotic stimuli that induce apoptosis in uninfected, neighbouring cells and that the redox state of the cell is a regulator in chlamydia-induced apoptosis.

  • 80. Strindhall, J
    et al.
    Lindgren, Per-Eric
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Medicinsk mikrobiologi.
    Löfgren, S
    Kihlström, Erik
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Clinical isolates of Staphylococcus aureus vary in ability to stimulate cytokine expression in human endothelial cells2005Ingår i: Scandinavian Journal of Immunology, ISSN 0300-9475, E-ISSN 1365-3083, Vol. 61, nr 1, s. 57-62Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Human umbilical vein endothelial cells (HUVEC) were infected for 24 h with 18 well-characterized Staphylococcus aureus isolates, and the supernatants from infected HUVEC were analysed for interleukin (IL)-1β, tumour necrosis factor-alpha, IL-6, IL-8, IL-10, IL-12p70, growth-related oncogene (GRO)-α, granulocyte macrophage colony-stimulating factor (GM-CSF) and regulated upon activation, normal T cell expressed and secreted (RANTES) by immunoassay. All staphylococcal isolates induced the expression of IL-6, IL-8, GRO-α, GM-CSF and RANTES. The magnitude of cytokine expression varied between isolates. Staphylococcus aureus inducing high expression of one of these cytokines also showed simultaneous high expression of the other four, indicating a common mechanism for the ability of individual S. aureus to induce expression of these cytokines. No direct correlation between cytokine expression and adhesion of S. aureus to HUVEC was observed, indicating that bacterial properties besides adhesion contribute to the activation of HUVEC.

  • 81.
    Strindhall, Jan
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Innate immune response in human endothelial cells: characterization and regulation of E-selectin, ICAM-I and cytokine expression and the role of Staphylococcus aureus2003Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Endothelial cells located on the vessel walls play an important role in inflammation. In response to cytokines or other inflammatory mediators such as microorganisms, endothelial cells express cell adhesion molecules such as E-selectin and ICAM-1 that are involved in binding circulating leukocytes to the endothelium. Since E-selectin is highly glycosylated one aim of this thesis was to investigate the role of N-glycosylation in E-selectin expression and function. Modification of glycosylation was obtained by culturing HUVEC in the presence of soluble glycosylation inhibitors. The result showed that E-selectin is highly glycosylated with N-Iinked, complex type oligosaccharides which were found to be important for the level of cell surface expression and turnover time. However, defect glycosylation did not affect its binding properties as revealed in a static cell-binding assay. Combinations of cytokines can produce a different expression of E-selectin. When TNF-α and IFN-γ were used to stimulate HUVEC an enhanced and prolonged expression of E-selectin was obtained compared to when HUVEC were stimulated with TNF-α alone. This effect could be blocked by monensin, which is an inhibitor of trans-Golgi network processing. This again indicated a role for posttranslational modification in regulation of E-selectin expression.

    Eighteen clinical isolates of Staphylococcus aureus were analysed for their ability to induce expression of E-selectin and ICAM-1 in HUVEC. Upregulation of adhesion molecules varied between isolates; 17 isolates induced expression of E-selectin and 13 of ICAM-1. Some isolates induced a significant expression of E-selectin without stimulation of ICAM-1, whereas the opposite was not found. Bacterial viability was required for induction of the adhesion molecules. There was no correlation between ability of S. aureus to induce expression of cell adhesion molecules and methicillin susceptibility, pulse field electrophoresis patterns, biochemical characteristics, phage typing or toxin production. Nine cytokines, IL-1ß, TNF-α, IL-6, IL-8, IL-10, IL-12p70, GRO-α, GM-CSF and RANTES, all of importance in the inflammatory process, were analysed in supernatants from HUVEC stimulated with eighteen isolates of S. aureus. All isolates induced IL-6, IL-8, GRO-α, GM-CSF and RANTES. Isolates of S. aureus inducing high expression of one of these cytokines also showed high expression of the other four, indicating a possible common mechanism for regulation of the expression of these cytokines. The ability of individual S. aureus isolates to induce expression of cytokines correlated with their ability to induce expression of ICAM-1 but not E-selectin in HUVEC. No correlation between cytokine profile and S. aureus production of enterotoxin A-D, TSST-1, cytotoxicity, PFGE- and phage pattern or susceptibility to methicillin was observed. Furthermore, culture filtrate from S. aureus induced expression of ICAM-1 and IL-8 in HUVEC. The component(s) from culture filtrates were heat-stable, had a molecular weight of about 100 kDa and the effect was independent of the presence of fetal calf serum in media.

    Delarbeten
    1. Role of N-linked glycosylation in expression of E-selectin on human endothelial cells
    Öppna denna publikation i ny flik eller fönster >>Role of N-linked glycosylation in expression of E-selectin on human endothelial cells
    1995 (Engelska)Ingår i: European Journal of Immunology, ISSN 0014-2980, E-ISSN 1521-4141, Vol. 25, nr 9, s. 2452-2459Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    E-selectin is a cytokine-inducible membrane glycoprotein capable of mediating adhesion of leukocytes to endothelial cells. It is highly glycosylated, containing 11 sites for N-linked glycosylation. N-Glycosylation of E-selectin was analyzed by endoglycosidase treatment. Analysis of immunoprecipitated E-selectin from human umbilical vein endothelial cells (HUVEC) by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate showed that E-selectin was completely resistant to endoglycosidase H, but sensitive to peptide N-glycanase F digestion. This suggested that all N-linked oligosaccharide chains were of the complex type. The role of N-linked glycosylation in surface expression and secretion of E-selectin was studied using interleukin-1-stimulated HUVEC, cultured in the presence of the soluble glycosylation inhibitors tunicamycin or castanospermine. Cell surface expression was analyzed by indirect flow cytometry. N-Glycosylation was blocked by tunicamycin, and resulted in a significantly reduced surface expression of E-selectin, whereas castanospermine only marginally reduced E-selectin expression. The deglycosylated forms of E-selectin were also found to be fully capable of mediating adhesion of HT-29 cells in vitro. In conclusion, these studies show that E-selectin is heavily glycosylated with complex type N-linked oligosaccharides and that N-glycosylation is important for expression of E-selectin on human endothelial cells.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-84628 (URN)10.1002/eji.1830250907 (DOI)
    Tillgänglig från: 2012-10-16 Skapad: 2012-10-16 Senast uppdaterad: 2017-12-07Bibliografiskt granskad
    2. Interferon-γ enhancement of E-selectin expression on endothelial cells is inhibited by monensin
    Öppna denna publikation i ny flik eller fönster >>Interferon-γ enhancement of E-selectin expression on endothelial cells is inhibited by monensin
    1997 (Engelska)Ingår i: Scandinavian Journal of Immunology, ISSN 0300-9475, E-ISSN 1365-3083, Vol. 46, nr 4, s. 338-343Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The expression of E-selectin reaches a maximum 4–6 h after stimulation of human umbilical vein endothelial cells (HUVEC) in vitro with tumour necrosis factor-α (TNF-α) and then declines to basal level within 24 h. If interferon-γ (IFN-γ) is added to the cell culture medium together with TNF-α the surface expression of E-selectin is augmented and prolonged in a synergistic way. The aim of the present study was to investigate if altered protein glycosylation could explain the IFN-γ induced persistent surface expression of E-selectin. SDS–PAGE analysis of HUVEC glycoproteins, metabolically radiolabelled in the carbohydrate portion, indicated that addition of IFN-γ produced an altered protein glycosylation. Lectin blot analysis using the Sambucus nigra agglutinin lectin also indicated differences in protein glycosylation when HUVEC were incubated with IFN-γ/TNF-α compared to TNF-α alone. The kinetics of surface expression of E-selectin were measured using a cell ELISA assay. When HUVEC were incubated with monensin, a potent inhibitor of late Golgi function, together with both TNF-α and IFN-γ, the additive effect of IFN-γ on E-selectin expression was almost abolished. Since monensin is known to affect glycosylation processing, this experiment suggested that the IFN-γ induced change in protein glycosylation might induce the prolonged surface expression of E-selectin. However, when HUVEC were cultured with IFN-γ/TNF-α in the presence of several different inhibitors of N-glycosylation processing, no significant effect of E-selectin expression was observed. Regulation of adhesion molecule expression after activation of endothelial cells is likely to play a pivotal role for the inflammatory response. Further studies are needed to understand the mechanisms underlying this regulation.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-84629 (URN)10.1046/j.1365-3083.1997.d01-135.x (DOI)
    Tillgänglig från: 2012-10-16 Skapad: 2012-10-16 Senast uppdaterad: 2017-12-07Bibliografiskt granskad
    3. Variations among clinical isolates of Staphylococcus aureus to induce expression of E-selectin and ICAM-1 in human endothelial cells
    Öppna denna publikation i ny flik eller fönster >>Variations among clinical isolates of Staphylococcus aureus to induce expression of E-selectin and ICAM-1 in human endothelial cells
    2002 (Engelska)Ingår i: FEMS Immunology and Medical Microbiology, ISSN 0928-8244, E-ISSN 1574-695X, Vol. 32, nr 3, s. 227-235Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Eighteen clinical isolates of Staphylococcus aureus, nine methicillin-sensitive and nine methicillin-resistant, were investigated for their ability to induce expression of E-selectin and ICAM-1 in human endothelial cells. Upregulation of adhesion molecules varied between isolates; 17 isolates induced expression of E-selectin and 13 of ICAM-1. Some isolates induced a significant expression of E-selectin without stimulation of ICAM-1, whereas the opposite was not found. Bacterial viability was required for induction of the adhesion molecules. The kinetics of ICAM-1 expression in S. aureus-infected cells differed from those stimulated with interleukin-1β (IL-1β). On the other hand, expression of E-selectin was very similar in S. aureus-infected and IL-1β-stimulated cells. There was no correlation between ability of S. aureus to induce expression of cell adhesion molecules, methicillin susceptibility, pulse field gel electrophoresis patterns, biochemical characteristics, phage typing and toxin production.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26508 (URN)10.1016/S0928-8244(01)00306-6 (DOI)11065 (Lokalt ID)11065 (Arkivnummer)11065 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    4. Cytokine expression in human endothelial cells stimulated with clinical isolates and culture filtrate of Staphylococcus aureus
    Öppna denna publikation i ny flik eller fönster >>Cytokine expression in human endothelial cells stimulated with clinical isolates and culture filtrate of Staphylococcus aureus
    (Engelska)Manuskript (preprint) (Övrigt vetenskapligt)
    Abstract [en]

    Human umbilical vein endothelial cells (HUVEC) were infected for 24 h with 18 well-characterized, noninvasive Staphylococcus aureus isolates and the supernatants were analyzed for IL-1ß, TNF-α, IL-6, IL-8, IL-10, IL-12p70, GRO-α, GM-CSF and RANTES by immuno assay. All isolates induced expression of IL-6, IL-8, GRO-α, GM-CSF, and RANTES. The highest concentrations were observed for IL-6, IL-8 and GRO-α, which reached levels close to that of HUVEC stimulated with LPS (0.1µg ml-1). The magnitude of cytokine expression varied between isolates. S. aureus inducing high expression of one of these cytokines also showed simultaneous high expression of the other four, indicating a common ability of individual S. aureus to induce high or low expression of these cytokines. IL-1ß, TNF-α, IL-10 and IL-12p70 were not upregulated by any of the S. aureus isolates. No correlation between cytokine profile and S. aureus production of enterotoxin A-D, TSST-1, cytotoxicity, PFGE and phage pattem or susceptibility to methicillin was observed. The ability of individual S. aureus to induce expression of cytokines correlated with their ability to upregulate ICAM-1, but not E-selectin, in HUVEC. Similarly, a heat-stable, high molecular weight component(s) from sterile culture filtrate of S. aureus upregulated expression of IL-8 and ICAM-1 in HUVEC.

    Our results show that individual clinical isolates of S. aureus vary in ability to directly stimulate human endothelial cells to upregulate cytoldnes that promote leukocyte recruitment and inflammation.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-84631 (URN)
    Tillgänglig från: 2012-10-16 Skapad: 2012-10-16 Senast uppdaterad: 2012-10-16Bibliografiskt granskad
  • 82.
    Strindhall, Jan
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Lindgren, Per-Eric
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Medicinsk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Löfgren, Sture
    Department of Microbiology, Hospital of Ryhov, Jönköping.
    Kihlström, Erik
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Cytokine expression in human endothelial cells stimulated with clinical isolates and culture filtrate of Staphylococcus aureusManuskript (preprint) (Övrigt vetenskapligt)
    Abstract [en]

    Human umbilical vein endothelial cells (HUVEC) were infected for 24 h with 18 well-characterized, noninvasive Staphylococcus aureus isolates and the supernatants were analyzed for IL-1ß, TNF-α, IL-6, IL-8, IL-10, IL-12p70, GRO-α, GM-CSF and RANTES by immuno assay. All isolates induced expression of IL-6, IL-8, GRO-α, GM-CSF, and RANTES. The highest concentrations were observed for IL-6, IL-8 and GRO-α, which reached levels close to that of HUVEC stimulated with LPS (0.1µg ml-1). The magnitude of cytokine expression varied between isolates. S. aureus inducing high expression of one of these cytokines also showed simultaneous high expression of the other four, indicating a common ability of individual S. aureus to induce high or low expression of these cytokines. IL-1ß, TNF-α, IL-10 and IL-12p70 were not upregulated by any of the S. aureus isolates. No correlation between cytokine profile and S. aureus production of enterotoxin A-D, TSST-1, cytotoxicity, PFGE and phage pattem or susceptibility to methicillin was observed. The ability of individual S. aureus to induce expression of cytokines correlated with their ability to upregulate ICAM-1, but not E-selectin, in HUVEC. Similarly, a heat-stable, high molecular weight component(s) from sterile culture filtrate of S. aureus upregulated expression of IL-8 and ICAM-1 in HUVEC.

    Our results show that individual clinical isolates of S. aureus vary in ability to directly stimulate human endothelial cells to upregulate cytoldnes that promote leukocyte recruitment and inflammation.

  • 83.
    Strindhall, Jan
    et al.
    Linköpings universitet, Institutionen för biomedicin och kirurgi, Klinisk kemi. Linköpings universitet, Hälsouniversitetet.
    Lindgren, Per-Eric
    Linköpings universitet, Institutionen för medicinsk teknik, Fysiologisk mätteknik. Linköpings universitet, Tekniska högskolan.
    Löfgren, Sture
    Department of Microbiology, Hospital of Ryhov, Jönköping, Sweden.
    Kihlström, Erik
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Variations among clinical isolates of Staphylococcus aureus to induce expression of E-selectin and ICAM-1 in human endothelial cells2002Ingår i: FEMS Immunology and Medical Microbiology, ISSN 0928-8244, E-ISSN 1574-695X, Vol. 32, nr 3, s. 227-235Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Eighteen clinical isolates of Staphylococcus aureus, nine methicillin-sensitive and nine methicillin-resistant, were investigated for their ability to induce expression of E-selectin and ICAM-1 in human endothelial cells. Upregulation of adhesion molecules varied between isolates; 17 isolates induced expression of E-selectin and 13 of ICAM-1. Some isolates induced a significant expression of E-selectin without stimulation of ICAM-1, whereas the opposite was not found. Bacterial viability was required for induction of the adhesion molecules. The kinetics of ICAM-1 expression in S. aureus-infected cells differed from those stimulated with interleukin-1β (IL-1β). On the other hand, expression of E-selectin was very similar in S. aureus-infected and IL-1β-stimulated cells. There was no correlation between ability of S. aureus to induce expression of cell adhesion molecules, methicillin susceptibility, pulse field gel electrophoresis patterns, biochemical characteristics, phage typing and toxin production.

  • 84.
    Sun, Yi-Qian
    et al.
    Linköpings universitet, Institutionen för biomedicin och kirurgi, Kirurgi. Linköpings universitet, Hälsouniversitetet.
    Monstein, Hans-Jürg
    Östergötlands Läns Landsting, LMÖ - Laboratoriemedicin i Östergötland.
    Nilsson, Lennart
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Petersson, Fredrik
    Pathology Research Department, Ryhov Hospital, Jönköping, Sweden.
    Borch, Kurt
    Linköpings universitet, Institutionen för biomedicin och kirurgi, Kirurgi. Linköpings universitet, Hälsouniversitetet.
    Profiling and identification of eubacteria in the stomach of Mongolian gerbils with and without Helicobacter pylori infection2003Ingår i: Helicobacter, ISSN 1083-4389, E-ISSN 1523-5378, Vol. 8, nr 2, s. 149-157Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Background. Mongolian gerbils are frequently used to study Helicobacter pylori-induced gastritis and its consequences. The presence of an indigenous bacterial flora with suppressive effect on H. pylori may cause difficulties with establishing this experimental model.

    Aim. The aim of the present study was to determine bacterial profiles in the stomach of Mongolian gerbils with and without (controls) H. pylori infection.

    Methods. Gastric tissue from H. pylori ATCC 43504 and CCUG 17874 infected and control animals were subjected to microbial culturing and histology. In addition, gastric mucosal samples from H. pylori ATCC 43504 infected and control animals were analyzed for bacterial profiling by temporal temperature gradient gel electrophoresis (TTGE), cloning and pyrosequencing of 16S rDNA variable V3 region derived PCR amplicons.

    Results. Oral administration of H. pylori ATCC 43504, but not CCUG 17874, induced colonization and gastric inflammation in the stomach of Mongolian gerbils. Temporal temperature gradient gel electrophoresis (TTGE) and partial 16S rDNA pyrosequencing revealed the presence of DNA representing a mixed bacterial flora in the stomach of both H. pylori ATCC 43504 infected and control animals. In both cases, lactobacilli appeared to be dominant.

    Conclusion. These findings suggest that indigenous bacteria, particularly lactobacilli, may have an impact on the colonization and growth of H. pylori strains in the stomach of Mongolian gerbils.

  • 85.
    Svensson, Erik
    et al.
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Ertzgaard, Per
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för nervsystem och rörelseorgan, Rehabiliteringsmedicin. Östergötlands Läns Landsting, Rekonstruktionscentrum, Rehabiliteringsmedicinska kliniken US.
    Forsum, Urban
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Bacteriuria in spinal cord injured patients with neurogenic bladder dysfunction2004Ingår i: Upsala Journal of Medical Sciences, ISSN 0300-9734, E-ISSN 2000-1967, Vol. 109, nr 1, s. 25-32Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The occurrence of bacteriuria in spinal cord injured patients with neurogenic bladder dysfunction who used clean intermittent catheterisation to empty their bladders was studied in order to examine cut-off concentration breakpoints for significant bacteriuria in this group of patients using procedures of the European Urinanalysis Guideline. 344 samples were cultured, yielding 285 isolates. Coagulase-negative staphylococci (27 %), Enterococci (25 %), Klebsiella spp (19 %), and Escherichia coli (12 %) were the most common findings. Bacteria grew at concentrations of 105-108 cfu/L, but only a few at 104 cfu/L. It is concluded that low bacterial concentrations in the urine (105 cfu/L) of patients with neurogenic bladder dysfunction who are on intermittent catheterisation might be as significant for bladder contamination with bacteria as a high bacterial concentration and can possibly be responsible for bladder infections.

  • 86.
    Söderquist, Bo
    et al.
    Örebro .
    Alriksson, Ingegerd
    Örebro.
    Källman, Jan
    Örebro.
    Kihlström, Erik
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    The influence of adhesive and invasive properties of Staphylococcus aureus defective in fibronectin-binding proteins on secretion of interleukin-6 by human endothelial cells2006Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 114, nr 2, s. 112-116Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Fibronectin-binding proteins (FnBP) are surface adhesins of Staphylococcus aureus documented to be virulence attributes in, for example, endovascular infections. By using mutants of S. aureus defective in the FnBPA and B genes we have investigated whether these adhesins affect cytokine expression in human umbilical vein endothelial cells (HUVEC). S. aureus expressing FnBPA and B adhered to and were internalized into HUVEC to a greater extent compared to mutants defective in expression of FnBP. Production and release of IL-6 was higher from endothelial cells infected with the parent FnBP-expressing strain compared to the FnBP-defective mutants. These results indicate that adhesion to and invasion of S. aureus into endothelial cells are important regulators of cytokine expression. © 2006 The Authors.

  • 87. Taha, Muhamed-Kheir
    et al.
    Olcén, Per
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Molecular genetic methods in diagnosis and direct characterization of acute bacterial central nervous system infections.2004Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 112, s. 753-770Artikel i tidskrift (Refereegranskat)
  • 88.
    Tegnell, Anders
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    Saeedi, Baharak
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Isaksson, Barbro
    Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Granfeldt, Hans
    Linköpings universitet, Institutionen för medicin och vård, Thoraxkirurgi. Linköpings universitet, Hälsouniversitetet.
    Öhman, Lena
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Linköpings universitet, Hälsouniversitetet.
    A clone of coagulase-negative staphylococci among patients with post-cardiac surgery infections2002Ingår i: Journal of Hospital Infection, ISSN 0195-6701, E-ISSN 1532-2939, Vol. 52, nr 1, s. 37-42Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Coagulase-negative staphylococci (CoNS) are important causes of hospital-acquired infections such as infections after cardiac surgery. Efforts to reduce these infections are hampered by the lack of knowledge concerning the epidemiology of CoNS in this setting. Forty strains of CoNS collected during the surgical revision of 27 patients operated on between 1997 and 2000 were analysed. Strains were also collected from the ambient air in the operating suite. Their pulsed-field gel electrophoresis (PFGE) characteristics and antibiotic resistance were analysed. Using PFGE 19 of 40 strains from 15 of 27 patients were shown to belong to one clone, and strains from this clone were also isolated from the ambient air. This clone had caused infections throughout the period. Antibiotic resistance did not correlate with PFGE patterns. Using PFGE one clone could be identified that caused 56% of the CoNS infections during this period. A strain from this clone was also found in the air of the operating suite suggesting the origin of the CoNS causing infections was the hospital environment.

  • 89.
    Tiveljung, Annika
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Identification and characterisation of bacteria based on 16S rDNA techniques with special reference to Helicobacter pylori in the gastro-intestinal tract2000Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    The overall aim of this study was to establish molecular techniques for the detection and identification of "difficult to grow" - bacteria in mixed bacterial populations in clinical samples without the need for culturing procedures.

    Material and Methods: Thirty-nine strains of Mobiluncus were used as a model system for phylogenetic classification of fastidious bacteria based on 16S ribosomal DNA (rDNA) sequences. To test the application of the 16S rDNA broad range PCR concept for detecting bacteria clinically, urine samples spiked with Chlamydia trachomatis elementary bodies and real urine test samples from 12 C. trachomatis positive and negative male volunteers were tested in a semiblind manner against routine procedures. Furthermore, gastric biopsy samples from 22 individuals (13 defined as having Helicobacter pylori-associated gastritis, and 9 defined as normal controls) were evaluated for the presence of Helicobacter 16S rDNA and the virulence genes cagA, vacA, and ureA. PCR products were also applied to temporal temperature gel electrophoresis (TTGE) gels for profiling the microbial flora. Finally, intestinal biopsies from 22 patients (11 diagnosed as Crohn's disease (CD), and 11 non-CD patients) were investigated using probes targeting potential pathogens that have been suggested to be involved in CD.

    Results: We were able to confirm the current species designation of Mobiluncus, although the results did not support the division of M. curtisii into subspecies. For the detection of C. trachomatis in urine, the in-house system was shown to be as sensitive as a commercially available PCR system. The search for Helicobacter pylori in gastric biopsies revealed the presence of Helicobacter DNA in 20 of 22 individuals. The molecular techniques were apparently too sensitive compared with routinely used techniques. TTGE revealed a complex microbial flora both in the normal control group and in the gastritis group, with dominance of Helicobacter in the gastritis group. The present results might lend support to the hypothesis that Helicobacter are indigenous biota of the human stomach. cagA was amplified in all Helicobacter positive specimens. None of the specimens in the control group carried a H. pylori type strain identical vacA genotype. ureA negative mutants were also found in this group. The 16S rDNA sequence data might also indicate phylogenetic heterogeneity. The mixed bacterial flora found in CD inflammatory lesions is consistent with the idea that the enteric microflora enters primary lesions, where secondary invaders may elicit chronic inflannnatory response.

    Conclusion: This thesis has demonstrated the usefulness of 16S rDNA based techniques for detection, identification, and characterisation of individual bacterial pathogens as well as for profiling of mixed bacterial flora in clinical specimens.

    Delarbeten
    1. Broad-range PCR amplification and DNA sequence analysis reveals variable motifs in 16S rRNA genes of Mobiluncus species
    Öppna denna publikation i ny flik eller fönster >>Broad-range PCR amplification and DNA sequence analysis reveals variable motifs in 16S rRNA genes of Mobiluncus species
    1995 (Engelska)Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 103, nr 7-8, s. 755-763Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Using DNA primers based on highly conserved regions of bacterial 16S ribosomal RNA genes, a technique was established for detection of Mobiluncus species by polymerase chain reaction (PCR) and hybridization analysis. Part of the 16S rRNA genes of Mobiluncus mulieris, Mobiluncus curtisii and uncharacterized Mobiluncus strains were analyzed by broad-range PCR amplification and direct DNA sequencing analysis. Sequence comparison of the partial 16S rRNA genes of Mobiluncus curtisii, Mobiluncus mulieris and atypical Mobiluncus strains studied indicated genus and species-specific motifs within the variable regions V3, V4 and V9 of 16S ribosomal DNAs. A Mobiluncus curtisii-specific primer, located within the variable region V3 of the 16S rRNA gene, was designed for Southern blot hybridization analysis of broad-range PCR products. Broad-range amplification combined with a M. curtisii-specific hybridization probe, Mob V3, distinguished between Mobiluncus curtisii, Mobiluncus mulieris, and atypical Mobiluncus strains.

    Nyckelord
    16S rRNA genes, PCR, DNA hybridization, bacterial vaginosis, Mobiluncus
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-79458 (URN)10.1111/j.1699-0463.1995.tb01434.x (DOI)
    Tillgänglig från: 2012-08-01 Skapad: 2012-08-01 Senast uppdaterad: 2017-12-07Bibliografiskt granskad
    2. Classification of the genus Mobiluncus based on comparative partial 16S rRNA gene analysis
    Öppna denna publikation i ny flik eller fönster >>Classification of the genus Mobiluncus based on comparative partial 16S rRNA gene analysis
    1996 (Engelska)Ingår i: International Journal of Systematic Bacteriology, ISSN 0020-7713, Vol. 46, nr 1, s. 332-336Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    On the basis of partial 16S rRNA gene sequences and the results of Southern blot analyses, we confirmed the division of the genus Mobiluncus into the species Mobiluncus curtisii and Mobiluncus mulieris. Division of M. curtisii into M. curtisii subsp. curtisii and M. curtisii subsp. holmesii was not supported by our data.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-79459 (URN)10.1099/00207713-46-1-332 (DOI)8573515 (PubMedID)
    Tillgänglig från: 2012-08-01 Skapad: 2012-08-01 Senast uppdaterad: 2017-12-07Bibliografiskt granskad
    3. Detection and identification of bacteria using in-house broad range 16S rDNA PCR amplification and genus-specific DNA hybridization probes, located within variable regions of 16S rRNA genes
    Öppna denna publikation i ny flik eller fönster >>Detection and identification of bacteria using in-house broad range 16S rDNA PCR amplification and genus-specific DNA hybridization probes, located within variable regions of 16S rRNA genes
    1996 (Engelska)Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 104, nr 1-6, s. 451-458Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Broad range PCR amplification and genus-specific 16S ribosomal DNA hybridization was used to demonstrate that Chlamydia, Helicobacter and Mobiluncus hybridization probes, located within variable regions V3, V4, and V9 of the 16S rDNA, specifically bound to the corresponding PCR product obtained from pure cultures of the three genera. The sensitivity of the assay was determined by analysis of C. trachomatis serially diluted in urine. The detection limit was 1–10 elementary bodies using a hybridization probe derived from the variable region V3 of the 16S rRNA gene. A PCR product was furthermore formed in urine specimens not containing C. trachomatis, showing amplification of chlamydia also in the presence of DNA from the resident urethral flora that competes for annealing sites. Analysis of a restricted number of male urine specimens using the C. trachomatis-specific probe showed complete agreement with culture and a commercially available PCR kit. Our method not only has the capacity to detect C. trachomatis in microbiologically mixed urine samples but also the potential advantage of identifying other bacterial pathogens from the same PCR product by varying the hybridization probes.

    Nyckelord
    16S rRNA genes, diagnostic PCR, Chlamydia trachomatis, Helicobacter pylori, Mobiluncus curtisii
    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-79460 (URN)10.1111/j.1699-0463.1996.tb00741.x (DOI)
    Tillgänglig från: 2012-08-01 Skapad: 2012-08-01 Senast uppdaterad: 2017-12-07Bibliografiskt granskad
    4. Identification of Helicobacter in gastric biopsies by PCR based on 16S rDNA sequences: a matter of little significance for the prediction of H. pylori-associated gastritis?
    Öppna denna publikation i ny flik eller fönster >>Identification of Helicobacter in gastric biopsies by PCR based on 16S rDNA sequences: a matter of little significance for the prediction of H. pylori-associated gastritis?
    Visa övriga...
    1998 (Engelska)Ingår i: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 47, nr 8, s. 695-704Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The aim of the present study was to correlate molecular evidence of the presence of Helicobacter pylori in gastric biopsy samples, based on analysis of 16S rDNA, vacuolating toxin (vacA), urease A (ureA) and cagA genes, with the clinical, histological and serological findings in patients with H. pylori-associated gastritis. Fresh biopsy samples were collected from the gastric antrum and corpus of 22 asymptomatic volunteers with or without H. pylori-associated gastritis. Total DNA was extracted from the biopsy material and subjected to 16S rDNA PCR amplification, Southern blotting and 16S rDNA sequence analysis of the PCR products. The vacA, ureA and cagA genes were characterised by PCR amplification and Southern blot analysis. Based on partial 16S rDNA sequence analysis, DNA belonging to the genus Helicobacter was detected in gastric biopsy samples from 20 of 22 subjects, including seven of nine histologically and serologically normal controls. Six of 20 partial 16S rDNA sequences revealed variations within variable regions V3 and V4 that deviated from those of the H. pylori type strain ATCC 4350T and, therefore, possibly represented other species of Helicobacter. VacA genes identical with those of the type strain were found predominantly in the subjects with H. pylori gastritis, and all the patients except one were found to be cagA-positive. There was no evidence of false positive PCR reactions. In conclusion, the PCR-based molecular typing methods used here were apparently too sensitive when applied to the detection of H. pylori in human gastric tissues. The lack of quantitative analysis makes them inappropriate as clinical tools for the diagnosis of H. pylori-associated gastritis, despite the fact that they provide a qualitative and sensitive tool for the detection and characterisation of H. pylori in the gastrointestinal tract.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-79462 (URN)10.1099/00222615-47-8-695 (DOI)9877190 (PubMedID)
    Tillgänglig från: 2012-08-01 Skapad: 2012-08-01 Senast uppdaterad: 2017-12-07Bibliografiskt granskad
    5. Profiling of bacterial flora in gastric biopsies from patients with Helicobacter pylori-associated gastritis and histologically normal control individuals by temperature gradient gel electrophoresis and 16S rDNA sequence analysis
    Öppna denna publikation i ny flik eller fönster >>Profiling of bacterial flora in gastric biopsies from patients with Helicobacter pylori-associated gastritis and histologically normal control individuals by temperature gradient gel electrophoresis and 16S rDNA sequence analysis
    Visa övriga...
    2000 (Engelska)Ingår i: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 49, nr 9, s. 817-822Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The aim of this study was to establish bacterial profiles in gastric biopsy specimens from patients with Helicobacter pylori-associated gastritis by means of temporal temperature gradient gel electrophoresis (TTGE) of PCR-amplified 16S rDNA fragments. Specimens from eight patients with asymptomatic gastritis and five histologically normal controls revealed a Helicobacter-specific band in the TTGE profile with increased amounts of Helicobacter-specific DNA in the biopsies from most of the gastritis patients. DNA from other genera including Enterococcus, Pseudomonas, Streptococcus, Staphylococcus and Stomatococcus was also found in the stomach. In the absence of gastric inflammation, Helicobacter spp. appeared to be part of a complex, presumably indigenous microbial flora found in the biopsy specimens from the stomach.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-24957 (URN)10966230 (PubMedID)9368 (Lokalt ID)9368 (Arkivnummer)9368 (OAI)
    Tillgänglig från: 2009-10-07 Skapad: 2009-10-07 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    6. Presence of eubacteria in biopsies from Crohn's disease inflammatory lesions as determined by 16S rRNA gene-based PCR
    Öppna denna publikation i ny flik eller fönster >>Presence of eubacteria in biopsies from Crohn's disease inflammatory lesions as determined by 16S rRNA gene-based PCR
    Visa övriga...
    1999 (Engelska)Ingår i: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 48, nr 3, s. 263-268Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The aim of this study was to search for putative microbial agents in Crohn's disease (CD) tissues by bacterial broad-range 16S rDNA PCR combined with genus- and species-specific DNA hybridisation analysis. Biopsies taken both surgically and endoscopically from the terminal ileum of 11 CD patients and 11 control patients were investigated. Significant amounts of eubacteria were demonstrated in biopsies taken endoscopically from both affected and unaffected individuals; the biopsies taken at surgery from control patients were negative. Three of five biopsies taken surgically from CD patients harboured Helicobacter spp.-, Mycobacterium paratuberculosis-, Listeria monocytogenes- and Escherichia coli-like 16S rDNA sequences. These findings show the importance of the sampling method chosen when combined with molecular typing of eubacteria in intestinal tissues. The mixed bacterial flora found in the surgical biopsies from CD patients supports the idea that the enteric microflora enters primary lesions where secondary bacterial colonisers may elicit a chronic inflammatory syndrome.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-25305 (URN)10.1099/00222615-48-3-263 (DOI)10334593 (PubMedID)9746 (Lokalt ID)9746 (Arkivnummer)9746 (OAI)
    Tillgänglig från: 2009-10-07 Skapad: 2009-10-07 Senast uppdaterad: 2017-12-13
  • 90.
    Tiveljung, Annika
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Backström, Jan
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Forsum, Urban
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet. Östergötlands Läns Landsting, Diagnostikcentrum, Klinisk mikrobiologi.
    Monstein, Hans-Jürg
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet. Östergötlands Läns Landsting, Diagnostikcentrum, Klinisk mikrobiologi.
    Broad-range PCR amplification and DNA sequence analysis reveals variable motifs in 16S rRNA genes of Mobiluncus species1995Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 103, nr 7-8, s. 755-763Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Using DNA primers based on highly conserved regions of bacterial 16S ribosomal RNA genes, a technique was established for detection of Mobiluncus species by polymerase chain reaction (PCR) and hybridization analysis. Part of the 16S rRNA genes of Mobiluncus mulieris, Mobiluncus curtisii and uncharacterized Mobiluncus strains were analyzed by broad-range PCR amplification and direct DNA sequencing analysis. Sequence comparison of the partial 16S rRNA genes of Mobiluncus curtisii, Mobiluncus mulieris and atypical Mobiluncus strains studied indicated genus and species-specific motifs within the variable regions V3, V4 and V9 of 16S ribosomal DNAs. A Mobiluncus curtisii-specific primer, located within the variable region V3 of the 16S rRNA gene, was designed for Southern blot hybridization analysis of broad-range PCR products. Broad-range amplification combined with a M. curtisii-specific hybridization probe, Mob V3, distinguished between Mobiluncus curtisii, Mobiluncus mulieris, and atypical Mobiluncus strains.

  • 91.
    Tiveljung, Annika
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Borch, Kurt
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Kirurgi. Linköpings universitet, Hälsouniversitetet. Östergötlands Läns Landsting, Centrum för kirurgi, ortopedi och cancervård, Kirurgiska kliniken i Östergötland.
    Jonasson, Jon
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet. Östergötlands Läns Landsting, Diagnostikcentrum, Klinisk mikrobiologi.
    Mårdh, Sven
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Cellbiologi. Linköpings universitet, Hälsouniversitetet.
    Petersson, Fredrik
    Ryhov Hospital, Jönköping.
    Monstein, Hans-Jürg
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet. Östergötlands Läns Landsting, Diagnostikcentrum, Klinisk mikrobiologi.
    Identification of Helicobacter in gastric biopsies by PCR based on 16S rDNA sequences: a matter of little significance for the prediction of H. pylori-associated gastritis?1998Ingår i: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 47, nr 8, s. 695-704Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The aim of the present study was to correlate molecular evidence of the presence of Helicobacter pylori in gastric biopsy samples, based on analysis of 16S rDNA, vacuolating toxin (vacA), urease A (ureA) and cagA genes, with the clinical, histological and serological findings in patients with H. pylori-associated gastritis. Fresh biopsy samples were collected from the gastric antrum and corpus of 22 asymptomatic volunteers with or without H. pylori-associated gastritis. Total DNA was extracted from the biopsy material and subjected to 16S rDNA PCR amplification, Southern blotting and 16S rDNA sequence analysis of the PCR products. The vacA, ureA and cagA genes were characterised by PCR amplification and Southern blot analysis. Based on partial 16S rDNA sequence analysis, DNA belonging to the genus Helicobacter was detected in gastric biopsy samples from 20 of 22 subjects, including seven of nine histologically and serologically normal controls. Six of 20 partial 16S rDNA sequences revealed variations within variable regions V3 and V4 that deviated from those of the H. pylori type strain ATCC 4350T and, therefore, possibly represented other species of Helicobacter. VacA genes identical with those of the type strain were found predominantly in the subjects with H. pylori gastritis, and all the patients except one were found to be cagA-positive. There was no evidence of false positive PCR reactions. In conclusion, the PCR-based molecular typing methods used here were apparently too sensitive when applied to the detection of H. pylori in human gastric tissues. The lack of quantitative analysis makes them inappropriate as clinical tools for the diagnosis of H. pylori-associated gastritis, despite the fact that they provide a qualitative and sensitive tool for the detection and characterisation of H. pylori in the gastrointestinal tract.

  • 92.
    Tiveljung, Annika
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Forsum, Urban
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet. Östergötlands Läns Landsting, Diagnostikcentrum, Klinisk mikrobiologi.
    Monstein, Hans-Jürg
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet. Östergötlands Läns Landsting, Diagnostikcentrum, Klinisk mikrobiologi.
    Classification of the genus Mobiluncus based on comparative partial 16S rRNA gene analysis1996Ingår i: International Journal of Systematic Bacteriology, ISSN 0020-7713, Vol. 46, nr 1, s. 332-336Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    On the basis of partial 16S rRNA gene sequences and the results of Southern blot analyses, we confirmed the division of the genus Mobiluncus into the species Mobiluncus curtisii and Mobiluncus mulieris. Division of M. curtisii into M. curtisii subsp. curtisii and M. curtisii subsp. holmesii was not supported by our data.

  • 93.
    Tiveljung, Annika
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Söderholm, Johan D.
    Linköpings universitet, Institutionen för biomedicin och kirurgi, Kirurgi. Linköpings universitet, Hälsouniversitetet.
    Olaison, Gunnar
    Linköpings universitet, Institutionen för biomedicin och kirurgi, Kirurgi. Linköpings universitet, Hälsouniversitetet.
    Jonasson, Jon
    Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Monstein, Hans-Jürg
    Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Presence of eubacteria in biopsies from Crohn's disease inflammatory lesions as determined by 16S rRNA gene-based PCR1999Ingår i: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 48, nr 3, s. 263-268Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The aim of this study was to search for putative microbial agents in Crohn's disease (CD) tissues by bacterial broad-range 16S rDNA PCR combined with genus- and species-specific DNA hybridisation analysis. Biopsies taken both surgically and endoscopically from the terminal ileum of 11 CD patients and 11 control patients were investigated. Significant amounts of eubacteria were demonstrated in biopsies taken endoscopically from both affected and unaffected individuals; the biopsies taken at surgery from control patients were negative. Three of five biopsies taken surgically from CD patients harboured Helicobacter spp.-, Mycobacterium paratuberculosis-, Listeria monocytogenes- and Escherichia coli-like 16S rDNA sequences. These findings show the importance of the sampling method chosen when combined with molecular typing of eubacteria in intestinal tissues. The mixed bacterial flora found in the surgical biopsies from CD patients supports the idea that the enteric microflora enters primary lesions where secondary bacterial colonisers may elicit a chronic inflammatory syndrome.

  • 94.
    Unemo, Magnus
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Genotypic and phenotypic characterisation of Neisseria gonorrhoeae2003Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [sv]

    Bakterien Neisseria gonorrhoeae (gonokocken) orsakar den sexuellt överförbara infektionen gonorré som med eller utan allvarliga komplikationer, som exempelvis infertilitet och utomkvedshavandeskap, är ett globalt folkhälsoproblem. Infektionen var tidigare mycket vanlig i flertalet länder, så även i Sverige där incidensen kulminerade år 1970 varefter den sjönk i stort sett varje år fram till och med år 1996. Från och med 1997 började infektionens incidens öka igen i Sverige, vilket också kunde noteras i flera andra länder. Framförallt orsakades den svenska ökningen av en ökad inhemsk smittspridning bland yngre heterosexuella kvinnor och män samt homosexuella män. En ökad resistens hos bakterien mot flertalet antibiotika är ett känt problem sedan decennier. De senaste åren har en nationell och internationell intensifiering skett av forskningen. Fokus ligger på effektiva diagnostiska tekniker, optimala karakteriseringsmetoder, epidemiologiska studier samt identifiering och övervakning av begynnande eller ökande antibiotikaresistens. Även patogenes, virulens och vaccinutveckling studeras.

    I denna avhandlings arbete I identifierades, bland svenska N. gonorrhoeae isolat, en hög prevalens av nedsatt känslighet eller resistens mot flertalet av de traditionella antibiotika för behandling av gonorré. En korrelation mellan nedsatt känslighet/resistens och geografisk smittort, framförallt Asien, kunde fastställas. Flera olika antibiotika för effektiv behandling finns dock fortfarande tillgängliga. I arbete I-V identifierades en stor genetisk heterogenitet inom och mellan olika serologiska varianter (serovarer) av N. gonorrhoeae stammar. Detta belyser behovet av att använda högdiskriminerande (DNA-baserade) metoder för epidemiologisk karakterisering av bakterien. Effektiva DNA-baserade molekylärgenetiska tekniker, som pulsfältgelelektrofores (PFGE) av genomiskt DNA efter klyvning med restriktionsenzym och sekvenseling av porB genen, adapterades, optimerades och evaluerades mot traditionella fenatypiska metoder, som epidemiologiska verktyg för svenska N. gonorrhoeae isolat. Båda metoderna visade en signifikant högre diskriminemnde förmåga, reproducerbarhet och objektivitet än traditionell karakterisering (arbete II & III). En jämförelse mellan serologisk och genetisk porB-baserad typning av N. gonorrhoeae visade på stora svårigheter att komplett identifiera de använda monoklonala antikropparnas antigena epitoper hos porB (arbete IV). I arbete IV & V utvecklades en bestämning av genetisk grupp (genogrupp) och genetisk variant (genovar) baserad på realtids-PCR av hela porB genen med efterföljande sekvensering i realtidsformat, med hjälp av pyrosequencing teknologi, av korta högvariabla segment av porB genen. Metoden är mycket snabb, högdiskriminerande, reproducerbar, objektiv samt har en hög kapacitet. Metoden skulle kunna ersätta alternativt komplettera den bristfalliga men internationellt etablerade och rutinmässigt använda serovarbestämningen av N. gonorrhoeae för att identifiera spridningen av individuella stammar i samhället, karakterisera isolat i misstänkta kluster av gonorréfall och för rutinmässig partnerspårning.

    Delarbeten
    1. One year of Neisseria gonorrhoeae isolates in Sweden: the prevalence study of antibiotic susceptibility shows relation to the geographic area of exposure
    Öppna denna publikation i ny flik eller fönster >>One year of Neisseria gonorrhoeae isolates in Sweden: the prevalence study of antibiotic susceptibility shows relation to the geographic area of exposure
    Visa övriga...
    2002 (Engelska)Ingår i: International Journal of STD and AIDS (London), ISSN 0956-4624, E-ISSN 1758-1052, Vol. 13, nr 2, s. 109-114Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The aim of this study was to compare epidemiological data with antibiotic susceptibility patterns, so as to characterize the risk of infection with a highly resistant Neisseria gonorrhoeae strain. N. gonorrhoeae strains isolated in Sweden from February 1998 through January 1999 were tested for antibiotic susceptibility. Epidemiological data were received from each clinician reporting a case of gonorrhoea and these data were linked to the N. gonorrhoeae strains. A total of 348 N. gonorrhoeae isolates, representing 89% of all Swedish cases diagnosed during the 12-month period, were tested for antibiotic susceptibility. Of all isolates, 24% were β-lactamase-producing, and 18% had decreased susceptibility to ciprofloxacin (MIC>0.064 mg/l). All isolates were fully susceptible to ceftriaxone and spectinomycin. More than 99% of the isolates were fully susceptible to azithromycin. The antibiotic susceptibility varied with the places where patients were exposed to infection. When exposed in Asia, 63% of the isolates showed reduced susceptibility to ciprofloxacin, compared with 0-8.5% of the isolates from patients exposed in other places (RR=8.5, P<0.001). Ciprofloxacin cannot be recommended as the first choice of treatment if the place of exposure was in Asia.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26456 (URN)10.1258/0956462021924730 (DOI)11004 (Lokalt ID)11004 (Arkivnummer)11004 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    2. Pulsed-field gel electrophoresis as an epidemiologic tool for Neisseria gonorrhoeae: Identification of clusters within serovars
    Öppna denna publikation i ny flik eller fönster >>Pulsed-field gel electrophoresis as an epidemiologic tool for Neisseria gonorrhoeae: Identification of clusters within serovars
    2002 (Engelska)Ingår i: Sexually Transmitted Diseases, ISSN 0148-5717, E-ISSN 1537-4521, Vol. 29, nr 1, s. 25-31Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Background: The increasing incidence of gonorrhea in Sweden in 1998 was due to mostly domestic cases. Among these, two core groups were identified: homosexual men with serovar IB-2 and young heterosexuals with serovar IB-3.

    Goals: To explore the genetic homogeneity/heterogeneity within the predominant serovars, IB-2 and IB-3, of Neisseria gonorrhoeae in Sweden by pulsed-field gel electrophoresis (PFGE) and to compare these results to epidemiologic information, as well as examine the genetic diversity within and between the 25 other represented serovars of N gonorrhoeae.

    Study Design: By PFGE, 237 N gonorrhoeae isolates were examined, and the results were compared with epidemiologic data for the IB-2 and IB-3 isolates.

    Results: In 79% of the domestic IB-2 cases involving homosexuals and 66% of the domestic IB-3 cases involving young heterosexuals, the isolates were genetically indistinguishable by PFGE. A high genetic diversity was identified within and between the 27 included serovars.

    Conclusions: Examination by means of PFGE indicated that one N gonorrhoeae clone each of the serovars IB-2 and IB-3 created the majority of the two core groups of domestic cases.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26457 (URN)11773875 (PubMedID)11005 (Lokalt ID)11005 (Arkivnummer)11005 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    3. Molecular epidemiology of Neisseria gonorrhoeae: sequence analysis of the porB gene confirms presence of two circulating strains
    Öppna denna publikation i ny flik eller fönster >>Molecular epidemiology of Neisseria gonorrhoeae: sequence analysis of the porB gene confirms presence of two circulating strains
    Visa övriga...
    2002 (Engelska)Ingår i: Journal of Clinical Microbiology, ISSN 0095-1137, E-ISSN 1098-660X, Vol. 40, nr 10, s. 3741-3749Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The phenotypic and genotypic characteristics of Neisseria gonorrhoeae strains fluctuate over time both locally and globally, and highly discriminative and precise characterization of the strains is essential. Conventional characterization of N. gonorrhoeae strains for epidemiological purposes is mostly based on phenotypic methods, which have some inherent limitations. In the present study sequence analysis of porB1b gene sequences was used for examination of the genetic relationships among N. gonorrhoeae strains. Substantial genetic heterogeneity was identified in the porB genes of serovar IB-2 isolates (8.1% of the nucleotide sites were polymorphic) and serovar IB-3 isolates (5.2% of the nucleotide sites were polymorphic) as well as between isolates of different serovars. The highest degree of diversity was identified in the gene segments encoding the surface-exposed loops of the mature PorB protein. Phylogenetic analysis of the porB1b gene sequences confirmed previous findings that have indicated the circulation of one N. gonorrhoeae strain each of serovar IB-2 and serovar IB-3 in the Swedish community. These strains caused the majority of the cases in two domestic core groups comprising homosexual men and young heterosexuals, respectively, and were also detected in other patients. The phylogenetic analyses of porB gene sequences in the present study showed congruence, but not complete identity, with previous results obtained by pulsed-field gel electrophoresis of the same isolates. In conclusion, porB gene sequencing can be used as a molecular epidemiological tool for examination of genetic relationships among emerging and circulating N. gonorrhoeae strains, as well as for confirmation or discrimination of clusters of gonorrhea cases.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26455 (URN)10.1128/​JCM.40.10.3741-3749.2002 (DOI)11003 (Lokalt ID)11003 (Arkivnummer)11003 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    4. Comparison of serologic and genetic porB-based typing of Neisseria gonorrhoeae: consequences for future characterization
    Öppna denna publikation i ny flik eller fönster >>Comparison of serologic and genetic porB-based typing of Neisseria gonorrhoeae: consequences for future characterization
    2003 (Engelska)Ingår i: Journal of Clinical Microbiology, ISSN 0095-1137, E-ISSN 1098-660X, Vol. 41, nr 9, s. 4141-4147Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    Due to temporal changes in the epidemiology of gonorrhea, a precise characterization of Neisseria gonorrhoeae is essential. In the present study genetic heterogeneity in the porB genes of N. gonorrhoeae was examined, and serovar determination was compared to porB gene sequencing. Among 108 N. gonorrhoeae isolates, phylogenetic analysis of the entire porB alleles (924 to 993 bp) identified 87 unique sequences. By analyzing only the four to six most heterogeneous porB gene regions (174 to 363 bp), 86 out of these 87 genetic variants were identified. Consequently, analysis of shorter highly variable regions of the porB gene generates high-level discriminatory ability as well as fast, objective, reproducible, and portable data for epidemiological characterization of N. gonorrhoeae. Regarding putative antigenic epitopes of PorB for Genetic Systems monoclonal antibodies (MAbs), some of the previous findings were confirmed, but new findings were also observed. For several of the MAbs, however, the precise amino acid residues of PorB critical for single-MAb reactivity were difficult to identify. In addition, repeated serovar determination of 108 N. gonorrhoeae isolates revealed discrepancies for 34 isolates, mostly due to nonreproducible reactivity with single MAbs. Thus, the prospects of a genetic typing system with congruent translation of the serovar determination seem to be limited. In conclusion, analysis of short highly variable regions of the porB gene could form the basis for a fast molecular epidemiological tool for the examination of emergence and transmission of N. gonorrhoeae strains within the community.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26458 (URN)10.1128/​JCM.41.9.4141-4147.2003 (DOI)11006 (Lokalt ID)11006 (Arkivnummer)11006 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13Bibliografiskt granskad
    5. Genetic variant (genovar) determination of Neisseria gonorrhoeae by pyrosequencing of highly polymorphic segments of the porB gene
    Öppna denna publikation i ny flik eller fönster >>Genetic variant (genovar) determination of Neisseria gonorrhoeae by pyrosequencing of highly polymorphic segments of the porB gene
    (Engelska)Manuskript (preprint) (Övrigt vetenskapligt)
    Abstract [en]

    For prevention and control of gonorrhea, an objective, highly discriminating, and reproducible molecular epidemiological characterization of Neisseria gonorrhoeae is essential. In the present study, in the pursuance of providing such qualities, pyrosequencing technology, a fast real-time DNA sequence analysis, was applied to six short, highly polymorphic porB gene segments, with subsequent genetic variant (genovar) determination of the bacterial isolates. The DNA sequencing templates were obtained by real-time PCR amplification, which also included fluorescence melting curve analysis of the entire porB gene in order to determine the genogroup (porB1a or porB1b allele) prior to pyrosequencing analysis. The PSQ 96 MA system used allowed rapid (approximately 1.5 h) determination of 96 sequences of 20 to 65 correct nucleotides each. The results were reproducible and mostly in concordance with the results of conventional Sanger dideoxy-sequencing, with the exception of shorter read lengths and some uncertainty in determining the correct number of identical nucleotides in homopolymeric segments. The number of sequence variants identified in each one of the six highly polymorphic segments of the porB1a and porB1b alleles (encoding surface exposed amino acid loops of the mature porB protein) ranged from 5 to 11 and 8 to 39, respectively. Among porB1a isolates (n=22) and porB1b isolates (n=65), 22 and 64 unique genovars were identified. All isolates were typeable. The current results provide evidence of a high discriminatory ability, practically the same as sequencing of the entire porB gene. In conclusion, the fast and high-throughput pyrosequencing technology can be used for molecular epidemiological characterization of N. gonorrhoeae.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-84322 (URN)
    Tillgänglig från: 2012-10-04 Skapad: 2012-10-04 Senast uppdaterad: 2012-10-04Bibliografiskt granskad
  • 95.
    Unemo, Magnus
    et al.
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Olcén, Per
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Jonasson, Jon
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Molekylär och immunologisk patologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk patologi och klinisk genetik.
    Fredlund, Hans
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Molecular typing of Neisseria gonorrhoeae isolates by pyrosequencing of highly polymorphic segments of the porB gene2004Ingår i: Journal of Clinical Microbiology, ISSN 0095-1137, E-ISSN 1098-660X, Vol. 42, nr 7, s. 2926-2934Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    For prevention and control of gonorrhea, an objective, highly discriminating, and reproducible molecular epidemiological characterization of Neisseria gonorrhoeae is essential. In the present study, in pursuance of providing such qualities, pyrosequencing technology, a fast real-time DNA sequence analysis, was applied to six short, highly polymorphic porB gene segments, with subsequent genetic variant (genovar) determination of the bacterial isolates. The sequencing templates were obtained by real-time PCR amplification, which also included fluorescence melting curve analysis of the entire porB gene in order to determine the genogroup (porB1a or porB1b allele) prior to pyrosequencing analysis. The PSQ 96 MA system used allowed rapid (in approximately 1.5 h) determination of 96 sequences of 20 to 65 correct nucleotides each. The results were reproducible and mostly in concordance with the results of conventional Sanger dideoxy sequencing, with the exception of shorter read lengths and some uncertainty in determining the correct number of identical nucleotides in homopolymeric segments. The number of sequence variants identified in each of the six highly polymorphic segments of the porB1a and porB1b alleles (encoding surface-exposed amino acid loops of the mature PorB protein) ranged from 5 to 11 and from 8 to 39, respectively. Among porB1a isolates (n = 22) and porB1b isolates (n = 65), 22 and 64 unique genovars, respectively, were identified. All isolates were typeable. The present results provide evidence of a high discriminatory ability, practically the same as that for sequencing of the entire porB gene. In conclusion, the fast and high-throughput pyrosequencing technology can be used for molecular epidemiological characterization of N. gonorrhoeae.

  • 96.
    Walther, Sten
    et al.
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för medicin och vård, Anestesiologi. Östergötlands Läns Landsting, Hjärtcentrum, Thorax-kärlkliniken.
    Agvald-Öhman, Christina
    Blomqvist, Hans
    Monnet, Dominique
    Nilsson, Lennart
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Hanberger, Håkan
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Infektionsmedicin. Östergötlands Läns Landsting, Medicincentrum, Infektionskliniken i Östergötland.
    Multiresistenta bakterier kan bli allt större hot på svenska IVA.2006Ingår i: Läkartidningen, ISSN 0023-7205, E-ISSN 1652-7518, Vol. 103, nr 49, s. 3930-3933Artikel i tidskrift (Övrigt vetenskapligt)
    Abstract [en]

      

  • 97.
    Westergren, Viveka
    et al.
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Forsum, Urban
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Östergötlands Läns Landsting, Laboratoriemedicinskt centrum, Klinisk mikrobiologi.
    Nosocomial sinusitis2005Ingår i: Nosocomial sinusitis: a unique subset of sinusitis / [ed] Stein M, Caplan ES., New York: Taylor and Francis , 2005, s. 319-356Kapitel i bok, del av antologi (Övrigt vetenskapligt)
    Abstract [en]

    PURPOSE OF REVIEW:

    Nosocomial sinusitis is a complication of critically ill patients that is frequently not considered as a cause of fever and infection. While this disease has been described in the literature there have been few recent citations on this subject. This brief review will familiarize the reader with the current state of the art with regard to diagnosis complications and treatment of this problem.

    RECENT FINDINGS:

    Nasal and oral tubes have been the prime inciting events. Other risk factors have been facial trauma, inability to mobilize the patient and prior sinus disease. Patients usually present while in the intensive care unit; and there are few signs that suggest sinusitis to the critical care team. A number of complications including direct extension to the brain, lung and blood stream, as well as sepsis and even death have been described. The diagnosis is usually made with the help of specific radiographs or computed tomography scans when these modalities are used. The microbiology is quite different than sinusitis in the community. Staphylococcus spp., Pseudomonas spp. and other nosocomial organisms are frequently isolated when specific cultures are obtained. Treatment usually consists of removal of the tubes mobilizing the patient and institution of broad-spectrum antibiotics aimed at the offending organisms.

    SUMMARY:

    Nosocomial sinusitis continues to be a major problem causing morbidity and occasionally mortality in critically ill patients. Recent findings have suggested that a careful search for this disease and appropriate treatment if found can decrease both morbidity, mortality and subsequent other nosocomial infections

  • 98. Wheeldon, T-U
    et al.
    Granström, M
    Hoang, TTH
    Phuncarg, DC
    Nilsson, Lennart
    Linköpings universitet, Hälsouniversitetet. Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi.
    Sörberg, M
    The importance of the level of metronidazole resistance for the success of Helicobacter pylori eradication2004Ingår i: Alimentary Pharmacology and Therapeutics, ISSN 0269-2813, E-ISSN 1365-2036, Vol. 19, nr 12, s. 1315-1321Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Aims: To evaluate the role of antibiotic susceptibility for the treatment outcome of proton pump inhibitor-dependent and independent Helicobacter pylori eradication regimens. Methods: In a placebo-controlled clinical study of peptic ulcer patients with H. pylori infection, patients were randomized to receive lansoprazole, clarithromycin and tinidazole twice-daily, clarithromycin and tinidazole once-daily with lansoprazole or with placebo. Helicobacter pylori status was assessed by culture and antibiotic susceptibility by E-test minimal inhibitory concentration (MIC) in 205 clinical isolates. Results: Primary resistance to clarithromycin and metronidazole was 1 and 76%, respectively. In metronidazole susceptible strains eradication rates were similar at > 90% for all treatment groups (P = 0.49). With low-level metronidazole resistance (4 μg/ mL < MIC < 256 μg/mL), eradication rates were similar at >75% (P = 0.80). The major difference was found at high-level metronidazole resistance (MIC ≥ 256 μg/mL) with 95%, 58% and 21% eradication in the lansoprazole, clarithromycin and tinidazole twice-daily, lansoprazole, clarithromycin and tinidazole once-daily and placebo, clarithromycin and tinidazole once-daily groups, respectively (P < 0.001). Conclusion: In the absence of antibiotic resistance, a once-daily therapy of only clarithromycin and tinidazole can achieve a high rate of H. pylori eradication. Such a combination could offer a simpler and cheaper treatment option for developing countries. The standard, twice-daily proton pump inhibitor-based triple therapy was shown to be efficient in H. pylori eradication even in the presence of high-level metronidazole resistance.

  • 99.
    Wolrath, Helen
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Amines and Bacterial Vaginosis2002Licentiatavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Bacterial Vaginosis (BY) is a common syndrome, with a prevalence of 10-30% in women of childbearing age. The decisive pathogenetic factoris thought to have a microbiological origin, but so far no specific bacteria have been implicated in causing BV. Instead, it appears that BV is accompanied by a shift in the normal lactobacilli flora to a mixed vaginal anaerobic flora. Vaginal fluid from women with BY has also been reported to contain various amines, and several techniques have been used to identify these amines.

    We developed a sensitive gas chromatographic and mass spectrometric (GC-MS) method to analyze amines related to BV together with quantification of the amines isobutylamine, phenethylamine, putrescine, cadaverine and tyramine. The aim of our investigation was to study if the amine content in vaginal fluids is quantitatively related to BV, diagnosed according to the Nugent scoring system. Our results show that the production of putrescine, cadaverine and tyramine is a property of BV, and that samples from healthy women do not include these amines.

    Using a sensitive gas chromatographic method, we also analyzed and quantified vaginal fluids with respect to trimethylamine (TMA), the amine responsible for the fishy odor in BV. In order to obtain a proper identification of BV, the vaginal fluid samples were Gram-stained and diagnosed according to two procedures. Our results show that regardless of the scoring method used for diagnosis, vaginal fluids from women with BV generally contain elevated amounts of TMA, while samples from healthy women do not.

    In conclusion, the presence of specific amines is clearly a prominent finding in women with BV, and these amines can thus be used as selective markers or diagnostic tools for the syndrome.

    Delarbeten
    1. Analysis of bacterial vaginosis-related amines in vaginal fluid by gas chromatography and mass spectrometry
    Öppna denna publikation i ny flik eller fönster >>Analysis of bacterial vaginosis-related amines in vaginal fluid by gas chromatography and mass spectrometry
    2001 (Engelska)Ingår i: Journal of Clinical Microbiology, ISSN 0095-1137, E-ISSN 1098-660X, Vol. 39, nr 11, s. 4026-4031Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The presence of various amines in vaginal fluid from women with malodorous vaginal discharge has been reported before. The investigations have used several techniques to identify the amines. However, an optimized quantification, together with a sensitive analysis method in connection with a diagnostic procedure for vaginal discharge, including the syndrome of bacterial vaginosis, as defined by the accepted “gold standard,” has not been done before. We now report a sensitive gas chromatographic and mass spectrometric method for identifying the amines isobutylamine, phenethylamine, putrescine, cadaverine, and tyramine in vaginal fluid. We used weighted samples of vaginal fluid to obtain a correct quantification. In addition, a proper diagnosis was obtained using Gram-stained smears of the vaginal fluid that were Nugent scored according to the method of Nugent et al. (R. P. Nugent et al., J. Clin. Microbiol., 29:297–301, 1991). We found that putrescine, cadaverine, and tyramine occurred in high concentrations in vaginal fluid from 24 women with Nugent scores between 7 and 10. These amines either were not found or were found only in very low concentrations in vaginal fluid from women with Nugent scores of 0 to 3. There is a strong correlation between bacterial vaginosis and the presence of putrescine, cadaverine, and tyramine in high concentrations in vaginal fluid.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-25913 (URN)10.1128/JCM.39.11.4026-4031.2001 (DOI)000171934200034 ()10355 (Lokalt ID)10355 (Arkivnummer)10355 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13
    2. Trimethylamine content in vaginal secretion and its relation to bacterial vaginosis
    Öppna denna publikation i ny flik eller fönster >>Trimethylamine content in vaginal secretion and its relation to bacterial vaginosis
    2002 (Engelska)Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 110, nr 11, s. 819-824Artikel i tidskrift (Refereegranskat) Published
    Abstract [en]

    The presence of a fishy odor emanating from women who present with a malodorous vaginal discharge is well known. The odor is due to bacterial reduction of trimethylamine oxide to trimethylamine (TMA) in vaginal secretion. The release of TMA from specimens of vaginal fluid following the addition of alkali is often used in making a clinical diagnosis of bacterial vaginosis (BV). We now report a sensitive gas chromatographic method for analysis and quantification of TMA in vaginal fluid in which weighed samples were used. In addition, a proper diagnosis of BV was obtained using Gram-stained smears of the vaginal fluid according to the method of Nugent et al. (R. P. Nugent et al., J Clin Microbiol 1991;29:297–301). We also diagnosed BV according to Hallén et al. (A. Hallén et al. Genitourin Med 1987;63:386–9). TMA was present in all women with a Nugent score between 7 and 10 and in almost all women diagnosed with BV according to the method of Hallén et al. TMA was not found or was only found in very low concentrations in vaginal fluid from women with Nugent scores of 0 to 3. TMA was also found in four women with a negative sniff test. It seems that high levels of TMA in samples of vaginal fluid are typical for BV regardless of the scoring method used for diagnosis. However, low levels of TMA, <5 μg/g vaginal fluid, do not always correlate with BV.

    Nationell ämneskategori
    Medicin och hälsovetenskap
    Identifikatorer
    urn:nbn:se:liu:diva-26449 (URN)10.1034/j.1600-0463.2002.1101108.x (DOI)000180804800008 ()10997 (Lokalt ID)10997 (Arkivnummer)10997 (OAI)
    Tillgänglig från: 2009-10-08 Skapad: 2009-10-08 Senast uppdaterad: 2017-12-13
  • 100.
    Wolrath, Helen
    et al.
    Linköpings universitet, Institutionen för molekylär och klinisk medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Borén, Hans
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Kemi. Linköpings universitet, Tekniska högskolan.
    Hallén, Anders
    Dept. of Dermatology and Venereology, University Hospital, Uppsala, Sweden.
    Forsum, Urban
    Linköpings universitet, Institutionen för klinisk och experimentell medicin, Klinisk mikrobiologi. Linköpings universitet, Hälsouniversitetet.
    Trimethylamine content in vaginal secretion and its relation to bacterial vaginosis2002Ingår i: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 110, nr 11, s. 819-824Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The presence of a fishy odor emanating from women who present with a malodorous vaginal discharge is well known. The odor is due to bacterial reduction of trimethylamine oxide to trimethylamine (TMA) in vaginal secretion. The release of TMA from specimens of vaginal fluid following the addition of alkali is often used in making a clinical diagnosis of bacterial vaginosis (BV). We now report a sensitive gas chromatographic method for analysis and quantification of TMA in vaginal fluid in which weighed samples were used. In addition, a proper diagnosis of BV was obtained using Gram-stained smears of the vaginal fluid according to the method of Nugent et al. (R. P. Nugent et al., J Clin Microbiol 1991;29:297–301). We also diagnosed BV according to Hallén et al. (A. Hallén et al. Genitourin Med 1987;63:386–9). TMA was present in all women with a Nugent score between 7 and 10 and in almost all women diagnosed with BV according to the method of Hallén et al. TMA was not found or was only found in very low concentrations in vaginal fluid from women with Nugent scores of 0 to 3. TMA was also found in four women with a negative sniff test. It seems that high levels of TMA in samples of vaginal fluid are typical for BV regardless of the scoring method used for diagnosis. However, low levels of TMA, <5 μg/g vaginal fluid, do not always correlate with BV.

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