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Identification ofcagA Tyrosine Phosphorylation DNA Motifs in Helicobacter pylori Isolates from Peptic Ulcer Patients by Novel PCR-Restriction Fragment Length Polymorphism and Real-Time Fluorescence PCR Assays

Title: Identification ofcagA Tyrosine Phosphorylation DNA Motifs in Helicobacter pylori Isolates from Peptic Ulcer Patients by Novel PCR-Restriction Fragment Length Polymorphism and Real-Time Fluorescence PCR Assays
Authors: Owen, Robert J.; Sharp, Sally I.; Chisholm, Stephanie A.; Rijpkema, Sjoerd
Source: Journal of Clinical Microbiology ; volume 41, issue 7, page 3112-3118 ; ISSN 0095-1137 1098-660X
Publisher Information: American Society for Microbiology
Publication Year: 2003
Description: Cag pathogenicity island-containing Helicobacter pylori (type I) induces signal transduction pathways resulting in tyrosine phosphorylation of proteins adjacent to the site of bacterial adhesion on host gastric epithelial cells. Conventional block PCR-restriction fragment length polymorphism (RFLP) and real-time LightCycler (LC) PCR hybridization assays, validated by direct sequencing, were designed to test for the presence of three nucleotide sequences corresponding to tyrosine phosphorylation motifs (TPMs) A, B, and C in 84 isolates of H. pylori type I from patients in England. Overall, the PCR assays demonstrated that one or more TPMs were present in 62 strains (75%). Motif A was common (71% of strains), whereas motifs B and C were rarer (8% of strains). Strains lacking a TPM were typically vacuolating cytotoxin genotype vacA m2. Motif A was widely distributed in relation to disease severity and was more commonly (but not significantly [ P = 0.071]) associated with gastric ulcer than with duodenal ulcer (86 versus 56%). The LC hybridization assay provided a rapid means of detecting all three motifs, but RFLP analysis was more specific for TPM-A. TPMs provide novel additional strain markers for defining cagA variation, including identification of RFLP types within TPM-A. The presence of a particular TPM was not of direct diagnostic value, either singly or in combination, but the higher proportion of TPM-A strains in gastric ulcer patients merits further investigation.
Document Type: article in journal/newspaper
Language: English
DOI: 10.1128/jcm.41.7.3112-3118.2003
DOI: 10.1128/JCM.41.7.3112-3118.2003
Availability: https://doi.org/10.1128/jcm.41.7.3112-3118.2003; https://journals.asm.org/doi/pdf/10.1128/JCM.41.7.3112-3118.2003
Rights: https://journals.asm.org/non-commercial-tdm-license
Accession Number: edsbas.61EF1E78
Database: BASE