Abstract
＂Helicobacter pylori＂ is thought to show strain diversity. The differences in the pathology of ＂H. pylori may＂ have to do with this diversity. We saw the use of polymerase chain reaction (PCR) to amplify the ureC in ＂H. pylori＂ and the examination of restriction fragment length polymorphism (RFLP) within the ureC region as a means to type clinical isolates of ＂H. pylori＂, where point mutations in the sequence diversity of the ureC gene can be found. Likewise, we saw the use of randomly amplified polymorphic DNA (RAPD) to type genotypic variation in the gene order on physical maps, mosaicism in conserved gene, non-conserved genes and extragenetic elements. In this study, H. pylori genomic DNA was prepared from strains isolated from 22 unrelated patients and examined by PCR-based typing of ＂H. pylori＂ isolates using RAPD analysis of genomic DNA and RFLP analysis of ureC gene with HindIII, EcoRI and Sau3A. From the restriction enzyme digestion of an 820-bp fragment from within the ureC gene in ＂H. pylori＂ strains isolated from the 22 patients, we found only one pattern with HindIII or EcoRI, but 6 different patterns with Sau3A. Nine strains (40.9%) were grouped RFLP pattern 1 with Sau3A digestion of ure C region. We found 21 different RAPD patterns of genomic DNA. The H. pylori RFLP patterns of ureC gene gave much simpler profiles than those produced by RAPD analysis of gemonic DNA. In summary, H. pylori isolates have great genetic diversity among strains, which can be exploited to type isolates. RAPD on the whole bacterial genome can accurately distinguish between isolates. RFLP analysis with Sau3A of the ureC gene also had a good discriminatory power and group isolates into clusters. Comparing the genetic diversity of H. pylori isolated from different patients, these methods yield fast and reliable results on the transmission and pathogenicity of ＂H. pylori＂.
Published Version
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