Rapid Automated Genotyping of CYP2C19 and Helicobacter pylori 23S rRNA Gene in Gastric Juice

Abstract

AIM: Genotypes of Helicobacter pylori (H. pylori) 23S rRNA gene and human CYP2C19 are predictive determinants of efficacy of triple therapy with proton pump inhibitor, clarithromycin, and amoxicillin for eradicating H. pylori. We intended to develop a rapid assay system to determine these genotypes. METHODS: We designed primer sets and specific probes for determining SNPs from adenine to guanine at positions 2142 or 2143 of H. pylori 23S rRNA gene and *2 and *3 alleles of CYP2C19, using the novel full-automated rapid genetic analyzer (GENECUBE®). The results were compared with those acquired by direct nucleotide sequencing of 50 gastric tissue samples. We also determined the presence of SNPs in 132 additional samples of patients’ gastric juices. RESULTS: The GENECUBE® genotyping results of the 23S rRNA gene and CYP2C19 from gastric tissue samples were in complete agreement with those for direct sequencing. The sensitivity, specificity, and validity of GENECUBE® results were all 100% when compared with clinical data. GENECUBE® genotyping required approximately 30 min for a single test. CONCLUSIONS: The GENECUBE®-based system can simultaneously measure genotypes of H. pylori 23S rRNA gene and human CYP2C19 from gastric tissues and juices rapidly, which can greatly aid the tailored H. pylori eradication therapies.