结核分枝杆菌耐多药性基因芯片联合检测

目的 利用基因芯片技术检测结核分枝杆菌(MTB)对链霉素(SM)、乙胺丁醇(EMB)、利福平(RFP)和异烟肼(INH)等4种抗生素的耐药性.方法 应用基因芯片法联合检测300株MTB临床分离株的SM、EMB、RFP和INH耐药性,并与BACTEC960法检测结果进行比较,将结果不相符的菌株进行测序,并比较结果.结果 基因芯片法测定300株MTB临床分离株,SM敏感188株、耐药112株,以rpsL43、88位aag突变为agg为主;EMB敏感251株,耐药49株,以embB 306位atg突变为gtg为主;RFP敏感211株,耐药89株,以ropB 516位aag突变为gag为主;INH敏感210株,90株,以KatG 315位gtg突变为gag为主;基因芯片法检测SM、EMB、RFP和INH耐药的敏感性、特异性分别为81.1%,99%;60.0%,100%;72.0%,98%;75.0%,99%;2种测定方法结果不相符菌株中,基因测序结果显示,基因芯片法检测点突变具有高度特异性.结论 基因芯片法联合检测MTB对SM、EMB、RFP和INH的耐药性,快速且特异性高,可作为MTB临床检测耐药性的快速筛选方法.

Detection of multi-drug resistant Mycobacterium tuberculosis by combined gene-chip and its clinical application

Objective To develop a new genechip for rapid detection of multidrug resistant Myco bacterium tuberculosis(M TB) to streptomy cin(SM),ethamtubol(EMB),rifampin(RFP) and/or isoniazid(INH).Methods The resistances of 300 MTB clinical isolates to SM,EMB,RFP and INH were detected with gene chip.Results Among the 300 MTB clinical isolates,188 isolate swere SM sensitive,112 were SM resistant with most of mutation of aag to agg on rpsL 43 and 88; 251 isolates were EM B sensitive and 49 were EMB resistant with the most of mutation of atg to gtg on embB 306; 251 isolates were RFP sensitive,49 resistant,and the mutation were mostly aag to gag on ropB 516; 210 isolates were INH sensitive,90 resistant,and the mutation were mostly gtg to gag on KatG 315.The sensitivity and specificity of the four antibiotic resistance were 81.1% and 99%,60.0% and 100%,72.0% and 98%,75.0% and 99%,respectively.For the isolates with different results by gene chip and conventional method,gene sequencing was applied,and the results showed that genechip for mutation detection had high specificity.Conclusion The detection for SM,EMB,RFP and INH resistance of MT B by combined gene chip is rapid and specific and could be applied to fast clinic screening.