高分辨率熔解曲线对痰液中结核分枝杆菌耐药性分析  

Analysis of drug resistance of Mycobacterium tuberculosis in sputum by high resolution melting curve

作　　者：姚欣[1] 黎彧利 朱艮苗[1] YAO Xin;LI Yuli;ZHU Genmiao(Department of Clinical Laboratory, People′s Hospital of Banan District, Chongqing 401320, China)

机构地区：[1]重庆市巴南区人民医院检验科,重庆401320

出　　处：《新疆医科大学学报》2019年第5期629-632,637,共5页Journal of Xinjiang Medical University

基　　金：国家高新技术研究发展项目(863计划)(2015AA020110)

摘　　要：目的利用高分辨率熔解曲线检测痰液中结核分枝杆菌的耐药性。方法取250例肺结核患者的痰样品进行涂片镜检和琼脂平板耐药性分析,通过实时PCR对耐药基因位点进行扩增。通过熔解曲线,将rpoB基因作为利福平抗性的生物标志物,将katG基因和inhA启动子区域作为异烟肼抗性的生物标志物。比较药敏试验的结果和熔解曲线分析的结果,以评估灵敏度、特异性、阳性预测值和阴性预测值的参数。结果高分辨率熔解曲线对利福平耐药的敏感性为90.3%,特异性为90.4%,阳性预测值为84.8%,阴性预测值为94.0%。对异烟肼的耐药率为90.2%,特异性为93.9%,阳性预测值为91.1%,阴性预测值为93.3%。多药耐药结核病的敏感性、特异性、阳性预测值和阴性预测值分别为89.9%、90.6%、78.5%和95.9%。结论高分辨率熔解曲线分析可用于痰标本中多药耐药结核的快速诊断。Objective To detect the drug resistance of Mycobacterium tuberculosis in sputum by high resolution melting curve. Methods A total of 250 sputum samples from patients with pulmonary tuberculosis were subjected to smear microscopy and agar plate resistance analysis. The resistance gene loci were amplified by real-time PCR. RpoB gene was used as a biomarker of rifampicin resistance, and katG gene and inhA promoter region were used as biomarkers of isoniazid resistance by melting curve analysis. The results of the susceptibility test and the results of the melting curve analysis were compared to evaluate parameters of sensitivity, specificity, positive predictive value, and negative predictive value. Results The sensitivity to rifampicin resistance was 90.3%, specificity was 90.4%, PPV was 84.8%, and NPV was 94.0%. The resistance rate to isoniazid was 90.2%, specificity was 93.9%, PPV was 91.1%, and NPV was 93.3%. The sensitivity, specificity, positive predictive value and negative predictive value of multidrug-resistant tuberculosis were 89.9%, 90.6%, 78.5% and 95.9%, respectively. Conclusion Melting curve analysis can be used for rapid diagnosis of multidrug-resistant tuberculosis in sputum samples.

关 键 词：高分辨率熔解曲线 结核病 耐药性 

分 类 号：R446.5[医药卫生—诊断学]