兰州大学学报（医学版）2025，Vol.51Issue(5)：10-15,6.DOI:10.13885/j.issn.1000-2812.2025.05.002
PCR-HRM检测幽门螺杆菌克拉霉素耐药性技术建立与评价
Establishment and evaluation of PCR-HRM technology for detecting clarithromycin resistance in Helicobacter pylori
摘要
Abstract
Objective To establish a polymerase chain reaction-high resolution melting(PCR-HRM)curve analysis for detecting clarithromycin resistance genes in Helicobacter pylori and evaluate its diagnostic abili-ty for clarithromycin-resistant phenotypes.Methods From September 2017 to December 2018,245 gastric mucosal tissue samples were randomly collected from patients undergoing gastroscopy at five hospitals in Gansu Province.Helicobacter pylori was isolated and cultured under microaerophilic conditions(5%O2,10%CO2,85%N2),yielding 116 clinical isolates.The clarithromycin resistance phenotype was determined using E-test.Helicobacter pylori DNA was extracted from gastric mucosal specimens,and PCR-HRM was per-formed using specific primers designed based on the 23S rRNA gene sequence.The consistency between PCR-HRM and E-test in detecting clarithromycin-resistant phenotypes was compared.Additionally,DNA amplification fragments of the target region from randomly selected strains were sequenced to validate the accuracy of PCR-HRM results.Results E-test revealed a clarithromycin resistance rate of 60.34%.PCR-HRM correctly identified 88.57%of resistant strains and 97.83%of susceptible strains.The sensitivity,speci-ficity,and negative likelihood ratio for detecting clarithromycin resistance were 87.14%,97.83%,and 0.131,respectively,with positive and negative predictive values of 98.39%and 83.33%.PCR-HRM showed high consistency with E-test results(Kappa=0.825,P<0.001).DNA sequencing showed that the identification results of PCR-HRM for the clarithromycin-resistant phenotype were completely consistent with the geno-typing results.Conclusion PCR-HRM is a rapid,simple,accurate and cost-effective method for detecting clarithromycin resistance in Helicobacter pylori,demonstrating high clinical application value.关键词
聚合酶链式反应/高分辨率熔解曲线分析/幽门螺杆菌/抗生素耐药性/克拉霉素Key words
polymerase chain reaction/high-resolution melting curve analysis/Helicobacter pylori/antibiotic resistance/clarithromycin分类
医药卫生引用本文复制引用
蒲素贵,尚琪,许慧梅,马雪妮,贠建蔚,张德奎..PCR-HRM检测幽门螺杆菌克拉霉素耐药性技术建立与评价[J].兰州大学学报（医学版）,2025,51(5):10-15,6.基金项目
甘肃省自然科学基金资助项目(22JR5RA960) （22JR5RA960）
甘肃省联合科研重大资助项目(23JRRA1501) （23JRRA1501）
甘肃省科技重大专项资助项目(24ZDFA006) （24ZDFA006）
