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Application of PCR amplicon sequencing using a single primer pair in PCR amplification to assess variations in Helicobacter pylori CagA EPIYA tyrosine phosphorylation motifs
Linköping University, Department of Clinical and Experimental Medicine, Clinical Microbiology. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Microbiology.
Linköping University, Department of Clinical and Experimental Medicine, Cell Biology. Linköping University, Faculty of Health Sciences.
Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Centre for Laboratory Medicine, Department of Clinical Microbiology.
Linköping University, Department of Clinical and Experimental Medicine, Surgery. Linköping University, Faculty of Health Sciences. Östergötlands Läns Landsting, Centre of Surgery and Oncology, Department of Surgery in Östergötland.
2010 (English)In: BMC Research Notes, ISSN 1756-0500, E-ISSN 1756-0500, Vol. 3, no 35Article in journal (Refereed) Published
Abstract [en]

Background

The presence of various EPIYA tyrosine phosphorylation motifs in the CagA protein of Helicobacter pylori has been suggested to contribute to pathogenesis in adults. In this study, a unique PCR assay and sequencing strategy was developed to establish the number and variation of cagA EPIYA motifs.

Findings

MDA-DNA derived from gastric biopsy specimens from eleven subjects with gastritis was used with M13- and T7- sequence-tagged primers for amplification of the cagA EPIYA motif region. Automated capillary electrophoresis using a high resolution kit and amplicon sequencing confirmed variations in the cagA EPIYA motif region. In nine cases, sequencing revealed the presence of AB, ABC, or ABCC (Western type) cagA EPIYA motif, respectively. In two cases, double cagA EPIYA motifs were detected (ABC/ABCC or ABC/AB), indicating the presence of two H. pylori strains in the same biopsy.

Conclusion

Automated capillary electrophoresis and amplicon sequencing using a single, M13- and T7-sequence-tagged primer pair in PCR amplification enabled a rapid molecular typing of cagA EPIYA motifs. Moreover, the techniques described allowed for a rapid detection of mixed H. pylori strains present in the same biopsy specimen.

Place, publisher, year, edition, pages
BioMed Central, 2010. Vol. 3, no 35
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:liu:diva-54559DOI: 10.1186/1756-0500-3-35PubMedID: 20181142OAI: oai:DiVA.org:liu-54559DiVA: diva2:305395
Note

Original Publication: Hans-Jurg Monstein, Anneli Karlsson, Anna Ryberg and Kurt Borch, Application of PCR amplicon sequencing using a single primer pair in PCR amplification to assess variations in Helicobacter pylori CagA EPIYA tyrosine phosphorylation motifs, 2010, BMC Research Notes, (3), 35, . http://dx.doi.org/10.1186/1756-0500-3-35 Licensee: BioMed Central http://www.biomedcentral.com/

Available from: 2010-03-23 Created: 2010-03-23 Last updated: 2017-12-12Bibliographically approved

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Monstein, Hans-JurgKarlsson, AnneliRyberg, AnnaBorch, Kurt

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Clinical MicrobiologyFaculty of Health SciencesDepartment of Clinical MicrobiologyCell BiologyDepartment of Clinical and Experimental MedicineSurgeryDepartment of Surgery in Östergötland
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