Determination of Helicobacter pylori resistance to clarithromycin in biopsy samples of gastric mucosa using TaqMan® MGB probes

Abstract

Objective. To study primary resistance of H. pylori to clarithromycin in residents of Gomel region by real-time polymerase chain reaction (PCR RT) using TaqMan® MGB probes.Materials and methods. The study included 184 patients diagnosed with gastritis and duodenitis, K29, median age 48.5 years (25% and 75% were 37 and 61 years old). According to the patients’ questionnaires, no clarithromycin-based eradication therapy was administered. To determine the resistance of H. pylori to clarithromycin, a PCR RV method using TagMan® MGB probes was used.Results. All 184 tested DNA samples were positive for the Rnase P gene (ICS) and were considered in further analysis (Ct, HEX 20.20-34.14). DNA from the cagH gene (Ct, FAM 21.26-33.04), indicating infection with the bacterium, was confirmed in 152 samples (82.6%). DNA from the 23SrRNA gene (point mutations A2142G and A2143G) was detected in 16 of 152 DNA samples 10.5 % (Ct, Hex 20.24-31.17). The positive control samples had characteristic curve growth in the corresponding detection channels; no curve growth was observed in the negative samples.Conclusion. The primary resistance of H. pylori to clarithromycin in the residents of Gomel region was 10.5%, and the use of triple first-line eradication therapy, including PPIs, amoxicillin and clarithromycin, as empirical in this region is consistent with the Maastricht III-VI recommendations and Decree of the Ministry of Health of the Republic of Belarus of 01.06.2017 № 54: clinical protocol “Diagnosis and treatment of patients with digestive diseases.” The use of PCR RT using TaqMan® MGB probes is justified to determine the resistance of H. pylori to clarithromycin, to prescribe individualized treatment and to evaluate the effectiveness of eradication regimens.