Polymerase chain reaction based analysis of the cytotoxin associated gene pathogenicity island of Helicobacter pylori from saliva: an approach for rapid molecular genotyping in relation to disease status.

BACKGROUND AND AIMS: The genetic composition of the intricate cytotoxin associated gene pathogenicity island (cag PAI) of Helicobacter pylori is known to significantly influence the outcome of the disease. Hence, analysis of complete cag PAI of H. pylori isolated from saliva would be of immense importance in standardizing saliva as a reliable non-invasive diagnostic specimen and also to evaluate the type of H. pylori infection. The aim of the present study was to analyze the genes of cag PAI of H. pylori for their presence and correlating them with the disease status of the patients. METHODS: One hundred and twenty patients (55 duodenal ulcer [DU], 25 gastric ulcer and 40 non-ulcer dyspepsia [NUD]) were investigated for the present study. Eight pairs of oligonucleotide primers (cagA1, cagA2, cagAP1, cagAP2, cagE, cagT, LEC1 and LEC2) of five different loci; cagA, cagA promoter region, cagE which represents cagI region, cagT and LEC representing cagII were used to detect the presence of the cag PAI genes by polymerase chain reaction. RESULTS: The comprehensive analysis of the genes constituting cag PAI showed almost equivalent prevalence of all the genes between both the study groups (ulcer and NUD) included. Little significant difference was found in the percentage distribution in both the clinical groups. cagE and cagT were found in a larger proportion of the ulcer group (92.5% and 96.2%) compared with the NUD group (77.5% and 85%), respectively. CONCLUSION: Saliva could be efficiently used as a non-invasive source for H. pylori and cagT might be an important locus of the cag PAI, thus greatly influencing the disease condition of the subjects.

The cag pathogenicity island of Helicobacter pylori is disrupted in the majority of patient isolates from different human populations.

The cag pathogenicity island (cag-PAI) is one of the major virulence determinants of Helicobacter pylori. The chromosomal integrity of this island or the lack thereof is speculated to play an important role in the progress of the gastroduodenal pathology caused by H. pylori. We determined the integrity of the cag-PAI by using specific flanking and internally anchored PCR primers to know the biogeographical distribution of strains carrying fully integral cag-PAI with proinflammatory behavior in vivo. Genotypes based on eight selected loci were studied in 335 isolates obtained from eight different geographic regions. The cag-PAI appeared to be disrupted in the majority of patient isolates throughout the world. Conservation of cag-PAI was highest in Japanese isolates (57.1%). However, only 18.6% of the Peruvian and 12% of the Indian isolates carried an intact cag-PAI. The integrity of cag-PAI in European and African strains was minimal. All 10 strains from Costa Rica had rearrangements. Overall, a majority of the strains of East Asian ancestry were found to have intact cag-PAI compared to strains of other descent. We also found that the cagE and cagT genes were less often rearranged (18%) than the cagA gene (27%). We attempted to relate cag-PAI rearrangement patterns to disease outcome. Deletion frequencies of cagA, cagE, and cagT genes were higher in benign cases than in isolates from severe ulcers and gastric cancer. Conversely, the cagA promoter and the left end of the cag-PAI were frequently rearranged or deleted in isolates linked to severe pathology. Analysis of the cag-PAI genotypes with a different biogeoclimatic history will contribute to our understanding of the pathogen-host interaction in health and disease.