三种分子方法检测结核分枝杆菌利福平耐药性的比较  

Comparison of three molecular assays for detection of rifampin resistance in Mycobacterium tuberculosis

作　　者：刘立宾[1] 王静[1] 李浩 岳永宁[1] 陈园园[1] 鲍志坚[2] 朱敏[2] Liu Libin;Wang Jing;Li Hao;Yue Yongning;Chen Yuanyuan;Bao Zhijian;Zhu Min(Department of Tuberculosis Laboratory,Zhejiang Chinese Medicine and Western Medicine Integrated Hospital,Hangzhou 310003,China;Department of Tuberculosis,Zhejiang Chinese Medicine and Western Medicine Integrated Hospital,Hangzhou 310003,China)

机构地区：[1]浙江省中西医结合医院结核实验室,杭州310003 [2]浙江省中西医结合医院结核科,杭州310003

出　　处：《中华临床感染病杂志》2020年第4期270-275,共6页Chinese Journal of Clinical Infectious Diseases

基　　金：杭州市科委主动设计项目(20162013A04)。

摘　　要：目的评估Xpert MTB/RIF、荧光PCR熔解曲线和基因芯片技术用于快速筛查利福平耐药结核病的临床价值。方法收集2016年9月3日至2019年8月11日在浙江省中西医结合医院收治的经Bactec MGIT 960液体培养药敏确诊为结核病,且具有Xpert MTB/RIF和基因芯片技术结果的150例患者临床资料,对所有患者分离培养菌株行荧光PCR熔解曲线检测。以Bactec MGIT 960液体培养药敏结果为金标准,分析Xpert MTB/RIF、荧光PCR熔解曲线和基因芯片技术对利福平耐药结核病的诊断效能。绘制受试者工作曲线(ROC)评估三种方法在利福平耐药结核病诊断中的价值。结果以Bactec MGIT 960药敏结果为金标准,Xpert MTB/RIF、荧光PCR熔解曲线和基因芯片技术检测利福平耐药的敏感度分别为88.89%(16/18)、94.44%(17/18)、88.89%(16/18),特异度分别为96.21%(127/132)、96.21%(127/132)、95.45%(126/132)。三种检测方法的敏感度和特异度比较,差异均无统计学意义(P>0.05)。三种分子方法检测利福平耐药与Bactec MGIT 960药敏结果的一致性Kappa值分别为0.794、0.827、0.770。ROC曲线分析显示,Xpert MTB/RIF、荧光PCR熔解曲线和基因芯片技术的ROC曲线下面积分别为0.926、0.953、0.922。三种检测方法对利福平耐药均具有较好的诊断价值(P<0.01),但三种方法比较,差异均无统计学意义(P>0.05)。三种方法检测利福平耐药与Bactec MGIT 960药敏不一致结果共8例。结论Xpert MTB/RIF、荧光PCR熔解曲线和基因芯片技术三种分子方法对利福平耐药性检测能力相当,均具有较高的敏感度和特异度,适合快速筛查利福平耐药结核病。Objective To evaluate the diagnostic effects of Xpert MTB/RIF,Fluorescence PCR melting curve and gene chip technology for rapid screening of rifampicin-resistant tuberculosis.Methods The clinical data of 150 patients diagnosed with tuberculosis by Bactec MGIT 960 liquid culture drug susceptibility in Zhejiang Chinese Medicine and Western Medicine Integrated Hospital from September 2016 to August 2019 were collected,including Xpert MTB/RIF and gene chip results.The isolated and cultured strains from patients were subjected to fluorescence PCR melting curve detection.Using Bactec MGIT 960 drug susceptibility results as the reference,the diagnostic efficacy of Xpert MTB/RIF,Fluorescence PCR melting curve and gene chip technology for rifampicin resistance were analyzed,and the receiver operating characteristic curve(ROC)was drawn for comparative analysis.Results Take Bactec MGIT 960 as the gold standard,the sensitivity of Xpert MTB/RIF,Fluorescence PCR melting curve and gene chip technology for rifampicin resistance were 88.89%(16/18),94.44%(17/18),88.89%(16/18)respectively;the specificity were 96.21%(127/132),96.21%(127/132),95.45%(126/132),respectively.There was no statistically significant difference in the sensitivity and specificity among the three detection methods(P>0.05).The Kappa values of the three molecular methods for detecting rifampicin resistance were 0.794,0.827 and 0.770,respectively.The three detection methods have good diagnostic value for rifampicin resistance(P<0.01),but there is no statistically significant difference between the three methods(P>0.05).There were 8 cases of inconsistent results between the three methods and Bactec MGIT 960 drug sensitivity.Conclusion Xpert MTB/RIF,Fluorescence PCR melting curve and gene chip technology have comparable ability to detect rifampicin resistance,all of these have high sensitivity and specificity for detecting rifampicin resistance and are suitable for rapid screening.

关 键 词：分枝杆菌 结核 利福平耐药 荧光PCR溶解曲线 Xpert MTB/RIF 基因芯片 ROC曲线 

分 类 号：R378.911[医药卫生—病原生物学]