DNA芯片快速检测结核分枝杆菌对链霉素和乙胺丁醇的耐药性方法的建立  

Rapid detection of M.tuberculosis resistance to SM and EMB by the Gene Chip method

作　　者：石小华[1] 景奉香[2] 胡忠义[3] 崔振玲[3] 赵建龙[2] 朱蕾[4] 

机构地区：[1]南京医科大学附属南京儿童医院内科,江苏南京210008 [2]中科院上海微系统与信息技术研究所,上海200050 [3]复旦大学附属中山医院呼吸内科,上海200032 [4]上海市肺科医院,上海200433

出　　处：《南京医科大学学报（自然科学版）》2009年第4期435-440,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家863计划资助项目(2002AA2Z2021)

摘　　要：目的:利用基因芯片技术检测结核分枝杆菌(MTB)对链霉素(SM)和乙胺丁醇(EMB)耐药性的技术平台建立。方法:应用基因芯片法测定160株MTB临床分离株的SM和EMB耐药性,并与BACTEC960法结果进行比较,分别对结果相符与不相符的菌株进行测序。结果:基因芯片法测定160株MTB临床分离株,结果显示SM敏感111株、耐药49株,以rpsL43、88位aag突变为agg为主。芯片检测结果EMB敏感142株、耐药18株,耐EMB的MTB以embB306位atg突变为gtg为主。基因芯片法检测链霉素、乙胺丁醇耐药的敏感性、特异性分别为80.0%、99%和60.0%、100%。两种测定方法结果不符的菌株中,采用基因测序法测定,结果显示基因芯片法检测点突变具有高度的特异性。结论:基因芯片法检测MTB对SM与EMB耐药性方法具有快速、简单,特异性高,准确性高,可作为MTB的SM与EMB耐药性快速筛选方法。Objective：To establish a method of rapidly detecting Mycobacterium tuberculosis resistance to streptomycin and ethamtubol by gene-chip. Methods：To detect the SM and EMB resistance of 160 clinical isolates of M.tuberculosis by Gene Chip, and compare them with the results of the BACTEC960 method. Then validate the concordance and discrepancy result of the two methods by DNA Sequencing method. Results：Of 160 strains of M. tuberculosis clinical isolates, 111 strains were SM-susceptible and 49 strains were SM-resistible with Gene Chip while 100 strains were SM-susceptible and 60 strains were SM-resistible with BACTEC 960 method; 142 strains were EMB-susceptible and 18 strains were EMB-resistible with Gene Chip while 130 strains were SM-susceptible and 30 strains were EMB-resistible with BACTEC 960 method. According to the result amended by the BACTEC 960 method, the sensitivity, specialty and veracity were 80.0%, 99.0%, 91.9% and 60.0%, 100% and 92.5% respectively. The most frequent mutant sites are rpsL 43, 88 aag→agg in relation to SM-resistible and embB 306 atg→gtg in relation to EMB-resistible. Conclusion： Gene Chip method could detect the SM and EMB resistance of MTB easily and rapidly. At the same time it showed higher specialty and sensitivity, and could be used for clinical detection of SM-resistant and EMB-resistant strains.

关 键 词：分枝杆菌 结核 药敏试验 基因芯片 链霉素 乙胺丁醇 

分 类 号：R378.911[医药卫生—病原生物学]