贵州省两个结核病监测点RFP和INH耐药基因检测与分析  

Detection and analysis of RFP and INH resistance genes at two monitoring sites of tuberculosis resistance in Guizhou Province

作　　者：黄俊飞 陈依江[1] 陈旭 郑雯琳 陈慧娟[1] 陈玮[1] 肖子宇 李世军[1] HUANG Jun-fei;CHEN Yi-jiang;CHEN Xu;ZHENG Wen-lin;CHEN Hui-juan;CHEN Wei;XIAO Zi-yu;LI Shi-jun(Guizhou Center for Disease Control and Prevention,Guiyang,China 550001;The Second Hospital Affiliated with Guizhou University of Traditional Chinese Medicine)

机构地区：[1]贵州省疾病预防控制中心,贵州贵阳550001 [2]贵州中医药大学第二附属医院

出　　处：《中国病原生物学杂志》2019年第11期1312-1315,1319,共5页Journal of Pathogen Biology

基　　金：贵州省科技支撑计划计划项目(黔科合支撑[2019]2822号);贵州省科学技术基金项目(黔科合基础[2019]1186);贵州省高层次创新型人才培养项目黔科合平台人才项目[2018]5606);贵州省院士工作站建设项目(黔科合平台人才[2018]5627)

摘　　要：目的应用分子生物学技术对贵州省威宁县和仁怀市2个结核病省级耐药监测点2018年确诊结核病患者感染结核分枝杆菌(MTB)RFP和INH耐药基因进行检测与分析,为结核病的防治提供参考数据。方法采用半巢式全自动实时荧光定量PCR结合PCR-线性杂交酶显色法对威宁县和仁怀市97例确诊结核病患者的痰培养物进行MTB核酸及利福平(RFP)和异烟肼(INH)耐药基因检测,分析结核病患者性别年龄构成。结果威宁县52份痰培养物样品经半巢式全自动实时荧光定量PCR检测出MTB核酸阳性49份,阳性率94.23%,RFP耐药基因阳性3份,RFP耐药率为6.12%。对RFP耐药基因阳性标本进行菌株分离培养和PCR-线性杂交酶显色检测,结果显示其中1株为RFP和INH双重耐药,双重耐药率为2.04%;仁怀市45份样品经半巢式全自动实时荧光定量PCR检测出MTB核酸阳性33份,阳性率73.33%。对RFP耐药基因阳性的2份标本进行菌株分离培养和PCR-线性杂交酶显色检测,结果显示RFP和INH耐药基因均阳性,双重耐药率为6.06%。患者性别构成分析结果显示,威宁县结核病患者男女比例为1.9∶1,仁怀市为1.6∶1,均为男多于女。患者年龄段分析显示,以青年及中年(18-65岁)多见,威宁县占86.53%(45/52),仁怀市占75.56%(34/45)。结论威宁县结核病患者感染MTB RFP耐药率6.12%,RFP和INH双重耐药率为2.04%;仁怀市结核病患者感染MTB RFP耐药率为6.06%,RFP和INH双重耐药率为6.06%。两地患者中均呈现男多于女,且以青年及中年占多数,结果可为当地结核病的防治提供参考依据。Objective To use molecular biology techniques to detect and analyze rifampicin(RFP)and isoniazid(INH)resistance genes of Mycobacterium tuberculosis in cases of tuberculosis in Weining and Renhuai,which are two provincial monitoring sites for tuberculosis resistance in Guizhou Province in 2018,in order to provide a reference for tuberculosis prevention and treatment.Methods Automated semi-nested real-time PCR and a PCR-line probe assay were used to detect M.tuberculosis and RFP and INH resistance genes in 97 cultured sputum samples from patients in Weining and Renhuai,Guizhou Province.The gender and age group of patients with tuberculosis were analyzed.Results Of the 52 samples from Weining,49 were positive for nucleic acids of M.tuberculosis at a rate of 94.23%.Three samples were resistant to RFP,for a rate of RFP resistance of 6.12%.RFP-resistant samples were isolated and cultured,and the PCR-line probe assay indicated that 1 sample was resistant to RFP and INH.The rate of resistance to both drugs was 2.04%.Of the 45 samples from Renhuai,33 were positive for nucleic acids of M.tuberculosis at a rate of 73.33%.Two were resistant to RFP.Both were resistant to RFP and INH at a rate of 6.06%.Patient information indicated that the ratio of male to female patients with tuberculosis in Weining was 1.9:1 while the ratio was 1.6:1 in Renhuai.Most of the patients with tuberculosis were younger or middle-aged(ages 18-65),accounting for 86.53%(45/52)of patients in Weining and 75.56%(34/45)of patients in Renhuai.Conclusion In 52 cultured sputum samples from Weining,RFP resistance was 6.12%and the rate of resistance to both drugs was 2.04%.In 45 samples from Renhuai,RFP resistance was 6.06%,and the rate of resistance to both drugs was 6.06%.There were more male than female patients in both Weining and Renhuai,and patients were mainly younger or middle-aged.These findings have provided a reference for tuberculosis prevention and treatment in Weining and Renhuai.

关 键 词：结核分枝杆菌 耐药 利福平 异烟肼 

分 类 号：R378.911[医药卫生—病原生物学]