荧光PCR熔解曲线法在检测临床标本结核分枝杆菌及其耐药性中的应用

Application of fluorescence PCR melting curve assay to detect mycobacterium tuberculosis and its drug-resistance in clinical specimens

目的评价荧光PCR熔解曲线法检测临床标本结核分枝杆菌及其对利福平和异烟肼敏感性的性能。方法收集重庆市公共卫生医疗救治中心2016年7月至2018年7月进行过荧光PCR熔解曲线法结核检测的病例,分别与结核菌培养和Genexpert比较检测结核菌的灵敏度,与比例法药敏比较药敏结果。结果在结核菌检测上,荧光PCR熔解曲线法阳性率与结核菌培养相近,低于Genexpert;与比例法药敏相比,检测利福平药敏的灵敏度为93.2%,特异度为80.2%,符合率为84.3%;检测异烟肼药敏的灵敏度为92.6%,特异度为86.3%,符合率为88.6%。结论荧光PCR熔解曲线法可以高灵敏地直接对临床样本快速检测结核菌及其耐药性。

Objective To evaluate the ability of fluorescence PCR melting curve assay in detection of mycobacterium tuberculosis in clinical samples.Methods The cases tested fluorescence quantitative PCR melting curve assay in Chongqing public health medical center during Jul.2016 to Jul.2018 were collected.The sensitivities of fluorescence PCR melting curve assay were compared with MTB culture and Genexpert respectively.The results of rifampin and isoniazid drug susceptibility of fluorescence PCR melting curve assay were compared with that of standard proportion method.Results The positive detection rate of MTB of fluorescence quantitative PCR melting curve assay was close to MTB culture,but lower than that of Genexpert;Compared with the proportion method,the sensitivity,specificity and concordance rates of rifampicin were 93.2%,80.2% and 84.3% respectively;The sensitivity,specificity and concordance rates of isoniazid were 92.6%,86.3% and 88.6% respectively.Conclusion Fluorescence quantitative PCR melting curve assay can be used to detect MTB and its drug-resistance directly in clinical samples with high sensitivity.