Abstract： AIM To investigate the antibiotic resistance of Helicobacter pylori (H. pylori) in Zhejiang, and to compare the consistency of different methods for antibiotic resistance detection. METHODS From June 2017 to September 2017, 127 H. pylori strains were isolated from gastric mucosa tissues of 305 patients who underwent gastroscopy. The sensitivity of these strains to six kinds of antibiotics was determined by the agar dilution method. The related gene mutations in 23S rRNA and gyrA were determined by quantitative realtime polymerase chain reaction (RT-PCR) and gene sequencing. RESULTS Of the 127 H. pylori strains isolated, 124 were resistant strains and three were sensitive strains. There was no strain that was resistant to amoxicillin, tetracycline, and furazolidone. The resistance rates to clarithromycin, levofloxacin, or metronidazole were 33.86％ (43/127), 44.88％ (57/127), and 91.34％ (116/127), respectively, and the resistance rates to the triple antibiotics was 33.86％(43/127). The main gene mutations associated with antibiotic resistance were 23S rRNA (A2143G) and gyrA(C261A/G), with mutation frequencies of 42.5％ (54/127) and 15％ (19/127), respectively. The drug resistance detected by RT-PCR method was consistent with that by sequencing method, but had low consistency with traditional culture method. CONCLUSION The antibiotic resistance rates of H. pylori to clarithromycin and levofloxacin in Zhejiang Province are remarkably high. In clinical treatment, it is necessary to test antibiotic resistance to choose proper antibiotics individually to improve the eradication efficiency.