不同方法检测利福平耐药结核病及MTB rpoB基因突变的对比分析  

Analysis of detection of rifampicin-resistant tuberculosis and rpoB gene mutation by using different methods

作　　者：林勇明[1] 黄晓伟 林淑芳[1] 魏淑贞[1] 林建[1] 赵永[1] LIN Yong-ming;HUANG Xiao-wei;LIN Shu-fang;WEI Shu-zhen;LIN Jian;ZHAO Yong(TB Control and Prevention Department of Fujian Provincial Center for Disease Control and Prevention ,Fuzhou 350000,China)

机构地区：[1]福建省疾病预防控制中心结核病防治科,福州,350000 [2]福建省泉州市疾病预防控制中心结核病防治科

出　　处：《结核病与肺部健康杂志》2019年第2期121-126,共6页Journal of Tuberculosis and Lung Health

基　　金：福建省卫生计生中青年骨干人才培养项目(2016-ZQN-21)

摘　　要：目的探讨不同方法检测利福平耐药结核病的效能,并分析利福平耐药MTB菌株ropB基因突变情况.方法搜集2017年福建省泉州市下属10个县(市、区)结核病定点医院登记的同时经固体比例法药物敏感性试验(简称“比例法药敏试验”)、核酸反向线性探针杂交技术(属于GenoType MTBDRplus方法,简称“MTBDRplus")与利福平耐药实时荧光定量核酸扩增检测技术(GeneXpertMTB/RIF,简称“Xpert技术”)等3种方法检测的511例确诊肺结核患者对利福平的耐药情况,并对3种方法检测结果不相同但至少1种结果为利福平耐药的66株MTB菌株行ropB基因测序,分析55株获得rpoB基因序列菌株的ropB基因突变情况.结果511例检测患者中,比例法药敏试验、MTBDRplus和Xpert检测到利福平的耐药率分别为38.16%(195/511)、40.51%(207/511)和45.99%(235/511),MTBDRplus技术检测的利福平耐药率与比例法差异无统计学意义(X^2=0.590,P=0.442),而Xpert技术检测利福平耐药率明显高于比例法(X^2=6.424,P=0.011).以比例法药敏试验检测结果为标准,Xpert技术和MTBDRplus检测MTB菌株对利福平耐药的敏感度、特异度、一致率分别为93.85%(183/195)和95.38%(186/195)、83.54%(264/316)和93.35%(295/316)、87.48%(447/511)和94.13%(481/511).获得rpoB基因序列的55株菌株中,47株(85.46%)ropB基因在371~566位点间的17个位置发生了36种类型的突变;其中13株(23.64%)发生碱基缺失或插入,7株(12.73%)发生点突变,27株(49.09%)发生联合突变.45株(81.82%,45/55)为比例法检测利福平敏感而分子药敏试验检测耐药[包括14株(25.45%,14/55)MTBDRplus与Xpert检测对利福平均耐药的菌株、30株(54.55%,30/55)MTBDRplus检测对利福平敏感而Xpert检测为耐药、1株(1.82%,1/55)MTBDRplus检测对利福平耐药而Xpert检测为敏感的菌株],8株(14.55%,8/55)比例法药敏试验对利福平耐药而MTBDRplus与Xpert检测均敏感的菌株.MTBDRplus与Xpert检测对利福平耐药结果相同的�Objective To explore the efficacy of different methods on detection of rifampicin-resistant tuberculosis (TB) and to analyze the situation of ropB gene mutation of rifampicin-resistant strains. Methods A total of 511 patients who had been notified as pulmonary TB in 10 county or district TB designated hospitals of Quanzhou City, Fujian Province in 2017, and received 3 kinds of tests as follows: drug susceptibility test with proportional method on solid media (proportional method), reverse linear probe hybridization (GenoType MTBDRplus method) and GeneXpert MTB/RIF (Xpert), were enrolled in this study and their situation of rifampicin resistance was collected. Among 511 TB patients, the ropB gene sequencing of 66 MTB strains with rifampicin resistance, which were detected by one of the 3 test methods at least, were conducted and the situation of the ropB gene mutations in 55 strains, which the sequence of rpoB gene were obtained, was analyzed. Results Among 511 strains tested, the rifampicin resistance rates detected by proportional method, MTBDRplus and Xpert were 3& 16%(195/511), 40. 51%(207/511) and 45.99%(235/511), respectively. There was no significant difference in rifampicin resistance rate between MTBDRplus technique and proportional method (X^2 =0. 590, P=0. 442), while it was significantly higher by Xpert technique than that of proportional method (X^2=6. 424, P=0. 011). When the test result of proportional method was regarded at the standard, the sensitivity, specificity and consistency rates of Xpert and MTBDRplus to rifampicin resistance were 93.85%( 183/195) and 95.39%( 186/195 ), 83.54%( 264/316) and 93.35%(295/316)? 87.48%(447/511) and 94. 13%(481/511), respectively. Among the 55 strains that their rpoB gene sequences were obtained, 36 types of mutation at 17 locations between 371 and 566 loci happened in the ropB gene of 47 strains (85. 46%): 13 strains (23. 64%) had base deletion or insertion, 7 strains (12. 73%) had point mutation and 27 strains (49.09%) had joint mutation. Forty-five strains (8

关 键 词：分枝杆菌 结核 利福平 抗药性 细菌 微生物敏感性试验 基因测定 

分 类 号：R52[医药卫生—内科学] R440[医药卫生—临床医学]