Detection of Helicobacter Pylori-Specific Genes in the Oral Yeast

Abstract

Introduction and Aims: Until today human stomach is the only recognized habitat of H. pylori. However, recruitment of DNA-based methods has made possible the detection of H. pylori in water and oral cavity, thus suggesting faecal-oral and oral-oral routes for transmission of H. pylori, respectively. In this study yeast has been proposed as a common vector for transmission of H. pylori. Thus designed primers were recruited to target 16S rDNA and cagA genes in the oral yeasts by PCR.

Materials and Methods: Eighteen yeasts were examined microscopically for the presence of bacterial-like bodies. DNAs were extracted from oral yeasts using phenol-chloroform method. Amplification conditions were optimized as 33 cycles and annealing temperatures of 63 |o|C for 16S rDNA and 51 |oC and 52 |oC for cagA gene which was targetted in two steps. DNAs of H. pylori and saccharomyces cerevisiae were used as controls. PCR products of two genes from One yeast and from H. pylori were cloned in pCAP and subsequently subcloned in pSK+ and sequenced.

Results: Bacterial-like bodies were observed in all oral yeasts. The amplified products of 16S rDNA from all oral yeasts were homologous in size with those of H. pylori. 15/18 (83%) yeasts contained cagA gene, homologous to H. pylori. CagA was not amplified from three yeasts and S. cerevisiae. Analysis of sequenced products of 16S rDNA and cagA from one oral yeast showed 98% homology with those of H. pylori.

Conclusions: The presence of H. pylori inside the yeast was indicated by light microscopy and PCR. It appears that yeasts which are abundant in nature and thrive the mucosal surfaces of human might serve as reservoirs and vehicles of H. pylori.