2种结核分枝杆菌耐药基因突变检测方法诊断利福平耐药结核病的评估研究  

Evaluation of two kinds of Mycobacterium tuberculosis drug resistance gene mutation detection methods for diagnosis of rifampicin-resistant tuberculosis

作　　者：薛建昌 梁冰锋[2] 吴海峰[1] 郑浩[3] 孙志平[1] 任哲 Xue Jianchang;Liang Bingfeng;Wu Haifeng;Zheng Hao;Sun Zhiping;Ren Zhe(Department of Laboratory,Hebei Chest Hospital,Shijiazhuang Hebei 050041,China;Department of Nursing,Hebei Women’s Vocational College,Shijiazhuang Hebei 050091,China;Department of Ultrasound,Hebei Chest Hospital,Shijiazhuang Hebei 050041,China)

机构地区：[1]河北省胸科医院检验科,河北石家庄050041 [2]河北女子职业技术学院护理系,河北石家庄050091 [3]河北省胸科医院超声室,河北石家庄050041

出　　处：《遵义医科大学学报》2020年第2期249-253,258,共6页Journal of Zunyi Medical University

基　　金：河北省医学科学重点课题(NO:20170424)。

摘　　要：目的评价荧光定量PCR探针熔解曲线法(简称Melt PCR法)和Gene Xpert MTB/RIF技术(简称Xpert法),用于临床涂阳标本诊断利福平耐药结核病(Rifampicin resistant tuberculosis,RR-TB)的检测效能及应用价值。方法随机收集2016年2月至2018年6月在河北省胸科医院就诊肺结核患者结核分枝杆菌(Mycobacterium tuberculosis,MTB)涂阳标本278例,应用传统的结核菌比例法药敏试验(对照检测方法)、Melt PCR法和Xpert法同时进行RR-TB情况的检测,分析Melt PCR法和Xpert法检测RR-TB的效能。结果3种方法同时对278例MTB阳性标本的利福平耐药进行检测,耐药检出结果差异无统计学意义(P>0.05)。与传统药敏试验结果相比,Melt PCR法和Xpert法的敏感度、特异度、阳性预测值、阴性预测值、准确度和约登指数分别为97.96%、98.25%、92.31%、99.56%、98.20%、96.21%和95.92%、97.38%、88.68%、99.11%、97.12%、93.30%,Kappa值分别为0.940和0.904(P<0.05),均为高度一致。3种方法在痰液中的利福平耐药均显著高于支气管肺泡灌洗液,且差异具有统计学意义(P<0.05)。不同带菌量样本的利福平耐药检出结果差异无统计学意义(P>0.05)。结论Melt PCR法和Xpert法用于临床涂阳标本中检测RR-TB具有相似的高灵敏度和特异度等检测性能,可结合实际需求在耐多药结核病防治中推广和应用。Objective To evaluate the efficacy and application value of fluorescence quantitative PCR probe melting curve(Melt PCR method)and Gene Xpert MTB/RIF(Xpert method)in the diagnosis of rifampicin resistant tuberculosis(RR-TB)in clinical smear positive specimens.Methods A total of 278 cases of Mycobacterium tuberculosis(MTB)smear samples from tuberculosis patients in Hebei Province Chest Hospital from February,2016 to June,2018 were randomly collected.RR-TB was detected by traditional tuberculosis proportional drug sensitivity test,Melt PCR method and Xpert method,and the efficacy of Melt PCR method and Xpert method in detecting RR-TB was analyzed.Results There was no significant difference in rifampicin resistance among 278 MTB positive specimens by the three methods.Compared with the results of traditional susceptibility test of tuberculosis,the sensitivity,specificity,positive predictive value,negative predictive value,accuracy and Yoden index of Melt PCR and Xpert were 97.96%,98.25%,92.31%,99.56%,98.20%,96.21%and 95.92%,97.38%,88.68%,99.11%,97.12%,93.30%.The Kappa values were 0.940 and 0.904,which were highly consistent.The drug resistance of rifampicin in sputum was significantly higher than that in bronchoalveolar lavage fluid.There was no significant difference in rifampicin resistance among different bacterial samples.Conclusion Melt PCR method and Xpert method have similar high sensitivity and specificity in the detection of RR-TB in clinical smear positive specimens,which could be popularized and applied in the prevention and treatment of multi-drug resistant tuberculosis according to the actual needs.

关 键 词：比例法药敏试验 荧光定量PCR探针熔解曲线法 Gene Xpert MTB/RIF技术 肺结核 利福平 耐药 

分 类 号：R378.911[医药卫生—病原生物学] R446.5[医药卫生—基础医学]