膜芯片技术对结核分枝杆菌耐药性检测的研究  

A study on membrane chip technique for drug resistance detection of Mycobacterium tuberculosis

作　　者：李芳芳[1] 唐曙明[1] 李爱敏[1] 侯沪[1] 刘彩艳[1] 

机构地区：[1]深圳市人民医院龙华分院检验科,广东深圳518000

出　　处：《国际检验医学杂志》2013年第21期2792-2794,共3页International Journal of Laboratory Medicine

基　　金：深圳市科技局资助项目(201102151)

摘　　要：目的验证膜芯片技术在结核分枝杆菌对利福平(RFP)、异烟肼(INH)、链霉素(SM)和乙胺丁醇(EMB)耐药性检测中的应用价值。方法收集393例临床标本(包括痰液、尿液、胸腔积液、腹水标本),根据结核分枝杆菌耐药位点的DNA序列,设计用于检测rpoB、katG、inhA、rpsL和embB基因常见突变类型的膜芯片,对上述临床标本进行检测,并将其与传统微生物敏感性试验结果进行比较,对不相符结果用聚合酶链反应产物直接测序(PCR-DS)进行验证。结果以常规培养法为金标准,膜芯片法扩增结核分枝杆菌的敏感性、特异性、准确性分别为90.4%、98.8%和95.7%,其对结核分枝杆菌RFP、INH、SM和EMB耐药性检测的敏感性分别为92.3%、83.3%、68.8%和83.3%;准确性分别为98.6%、97.3%、96.6%、和98.5%;特异性均达到100.0%。结论膜芯片法检测结核分枝杆菌耐药性具有较高的敏感性、特异性和准确性,可作为常规微生物敏感性试验的补充。Objective To verify the application value of membrane chip technique in detecting resistance to rifampicin（RFP）, isoniazid（INH） ,streptomycin（SM） and ethambutol（EMB） in Mycobacterium tuberculosis. Methods 393 clinical specimens（including sputum,urine, pleurai effusion and ascites specimens） were collected. Membrane chip were designed to detect the common mu tation types of rpoB,katG,inhA,rpsL and embg genes,according to DNA sequence of drug resistance loci of Mycobacterium tuberculosis. The clinical specimens above were detected and the results were compared with those of traditional microbial sensitivity tests,and those which did not match each other were subjected to polymerase chain reaction-direct sequencing（PCR-DS） for verification. Results Conventional cluture method were served as golden standard,the sensitivity,specificity and accuracy of membrane chip method which used for amplifying Mycobacterium tuberculosis were 90.4 %, 98.8 % and 95.7 % , respectively. Its sensitivity for detecting drug resistance of Mycobacteriurn tuberculosis to RFP,INH,SM and EMB were 92.3% ,83.3% ,68.8% and 83.3% ,respectively,its accuracy were 98.6%, 97. 3%,96. 6% and 98.5%, respectively, and the specificity were all 100. 0%. Conclusion Membrane chip method shows higher sensitivity,specificity and accuracy for detecting drug resistance of Mycobacterium tuberculosis and may be used as a supplement of routine microbial sensitivity test.

关 键 词：分枝杆菌 结核 抗药性 微生物敏感性试验 膜芯片 

分 类 号：R440[医药卫生—诊断学]