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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.

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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