In vitro model of production of antibodies; a new approach to reveal the presence of key bacteria in polymicrobial environments
2016 (English)In: BMC Microbiology, ISSN 1471-2180, E-ISSN 1471-2180, Vol. 16, 209Article in journal (Refereed) Published
Background: There is a rapid emergence of multiple resistant gram-negative bacteria due to overuse of antibiotics in the treatment of infections. Biofilms consist of polymicrobial communities that survive the hosts defense system. The key bacteria in biofilms are slow growing and support an attachment and rapid growth of other microorganisms. Current antimicrobial strategies often fail due to poor diagnosis of key pathogens in biofilms. The study aims to develop anti-bacterial human antibodies in vitro from patients who had recently undergone a systemic infection by pathogenic bacteria and to use these antibodies as a tool for detecting bacteria in biofilms. Methods: Lymphocytes were separated from whole blood of patients (n = 10) and stimulated with heat-killed bacteria to produce antibodies in vitro. The specificity of antibodies in recognizing the bacteria against which they were directed was evaluated by surface plasmon resonance system (SPR) and electron microscopy. The ulcer secretions from patients with chronic and acute leg ulcers and healthy controls were analyzed by the SPR system and the results were compared with culture studies. Results: The produced antibodies recognized bacteria with high sensitivity (SPR). The antibodies against Enterococcus fecalis bound specifically to the microorganism in a bacterial co-culture that was visualized by electron microscopy. Conclusion: In the present work, a method for producing specific antibodies against bacteria is introduced to recognize bacterial components in body fluids of patients suffering from pathogenic biofilms. This diagnostic technique may be most useful in clinical microbiology and in the choice of antibiotics in the treatment of serious infections.
Place, publisher, year, edition, pages
BIOMED CENTRAL LTD , 2016. Vol. 16, 209
Enterococcus; Biofilms; Antibodies; Detection; Electron microscopy; Lymphocytes; Surface plasmon resonance
IdentifiersURN: urn:nbn:se:liu:diva-132539DOI: 10.1186/s12866-016-0821-5ISI: 000383425300002PubMedID: 27612600OAI: oai:DiVA.org:liu-132539DiVA: diva2:1046375
Funding Agencies|PEAS Institut; FORSS [F2004-212]; ALF grants [LIO-206241]2016-11-142016-11-132016-12-02