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Determination of Helicobacter pylori vacA allelic types by single‐step multiplex PCR
Author(s) -
Chisholm S.A.,
Teare E.L.,
Patel B.,
Owen R.J.
Publication year - 2002
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2002.01129.x
Subject(s) - multiplex polymerase chain reaction , helicobacter pylori , biology , multiplex , allele , polymerase chain reaction , genetics , microbiology and biotechnology , bacteria , gene
Aims: To develop and evaluate a novel multiplex PCR assay that enables definition ofHelicobacter pylori vacA allelic type in a single reaction. Methods and Results: Application of the one‐step system to DNA extracts from 22 cultures of knownvacAgenotype demonstrated that it was highly accurate. Analysis of 15 matched gastric biopsy/culture pairs generated exactly correlating genotype profiles.vacAgenotypes were determined from an additional 62/70 gastric biopsies from dyspeptic patients of knownH. pylori positive status by the one‐step assay, compared with 63/70 by the original two‐reaction test. Types s1/m1, s1/m2 and s2/m2 were identified in 51·9%, 31·2% and 16·9% of biopsies, respectively. Conclusions: The multiplex PCR system developed enables rapid one‐stepvacA genotyping that is accurate, easy to interpret and more economical than the alternative multiple‐reaction tests. Application of this system to gastric biopsies from patients in South‐east England demonstrated that s1/m1 was the most common genotype, while s1/m2 and s2/m2 were less prevalent. Significance and Impact of the Study: This simple one‐step system can be applied direct to antral gastric biopsies without the need for culture, thereby facilitating rapid surveillance ofvacA genotype in relation to geographical location and disease status.