High-resolution melting curve analysis of levofloxacin-resistant gyrA gene mutation in Helicobacter pylori

• DOI: 10.3760/cma.j.issn.1674-1927.2018.02.002
• Publication Date: 2018-04-15
• Authors: Lipei Wu; Yugui Zhou; Wei Zhang; Kang Xu
• Keywords: High Resolution Melt; Helicobacter; Melting curve analysis

Objective To optimize and establish a PCR-high resolution melting curve (HRM) for analysis of levofloxacin-resistant gyrA gene mutation in Helicobacter pylori, to provide evidence clinical selection individualized pylori eradication regimens.

Methods Gastric mucosal tissues were collected from 45 patients with peptic ulcer who underwent gastroendoscopy at the Department Gastroenterology, Dongtai Hospital Affiliated Nantong University between May 2013 October 2013. After culture under microaerobic condition (5% O2, 10% CO2, 85% N2) , 23 isolates strains obtained subjected levofloxacin susceptibility test by E-test. Total DNA was extracted gastric mucosa these isolates. Partial fragments amplified PCR then sequence performed. GyrA mutations analyzed PCR-HRM.

Results The rate resistance 47.8% (11/23) . mutated 10 drug-resistant strains, presenting chiefly as C261A, G271T A272G+ G514A. There no 9.1% (1/11) resistant all sensitive strains. The specificity reproducibility PCR-HRM method 100%, minimum detection limit 102 copies/μl.

Conclusion The may accurately rapidly detect pylori.

Key words: Helicobacter pylori; Drug resistance; Levofloxacin; Gene; Mutation; High