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Abstract

We previously identified four potential putative gastroduodenal disease fragments by using the interspersed repetitive extragenic palindromic DNA sequence based PCR (REP-PCR) technique. We investigated these fragments with regard to their disease specificity. The putative disease-specific REP-PCR fragments were cloned, mapped by restriction enzymes, cross-hybridized, and confirmed by Southern hybridization. The four fragments were also used as probes against REP-PCR amplicons from H. pylori isolates obtained from gastritis (N = 20), duodenal ulcer (N = 30), and gastric cancer patients (N = 30). Three of these fragments (1.4- and 0.76-kb for gastritis; 1.35 kb for duodenal ulcer) were amplified without any discrimination between any disease-specific H. pylori isolates. However, amplification following hybridization with the fourth 0.81-kb fragment was observed only from gastritis (60%) and duodenal ulcer (52%) but with none (0%) of gastric cancer patients. Nucleotide sequence analysis of the 0.81-kb fragment revealed that it was an open reading frame of the hypothetical protein HP0373 matched to the position of 380,966 to 383,068 nucleotides of the H. pylori complete genome sequence. Hence, the REP-PCR sequence was not a extragenic palindromic DNA sequence. The hypothetical protein was also present in all the tested isolates. The REP-PCR fingerprinting technique is useful to differentiate disease-specific H. pylori strains based on the interspersed repetitive extragenic palindromic DNA sequences; however, it may not be useful to identify disease-specific virulence determinant(s) without being confirmed by DNA sequence analysis and functional studies.

CRISPR-like sequences in Helicobacter pylori and application in genotyping

“…In studies of H. pylori , biotyping, serotyping and hemagglutinin typing, have been reported to possess low discriminatory power index (DI) compared to genotyping such as RAPD [ 14 ], pulse field gel electrophoresis (PFGE) [ 15 ], restriction fragment length polymorphism-PCR (RFLP-PCR) [ 16 ], and repetitive extragenic palindromic PCR fingerprinting (REP-PCR) [ 17 ]. PFGE is not widely used for H. pylori because inter-patient variation is rare in the fingerprints obtained [ 18 ].…”

A M r 34,000 proinflammatory outer membrane protein ( oipA ) of Helicobacter pylori

“…Inactivation of the genes was confirmed by PCR amplification followed by Southern blot hybridization. Isogenic galE mutant was constructed as previously described (13).…”

A Single Bifunctional UDP-GlcNAc/Glc 4-Epimerase Supports the Synthesis of Three Cell Surface Glycoconjugates in Campylobacter jejuni

“…The insertional inactivation of galE in N. meningitidis MC58 and N. gonorrhoeae MS11 had a similar effect on the LOS (50,51). The LPS was truncated in galE mutants of Haemophilus influenzae (52,53) and Helicobacter pylori (54) as well as in a gne (lse) mutant of Y. enterocolitica of serotype O:8 (19). The O-antigen was lost in a gne mutant of E. coli strains of serotypes O55 and O157 (36).…”