PCR-SSCP法检测结核分枝杆菌耐药基因突变分析  

Analysis on the mutation detection of drug resistance genes of mycobacterium tuberculosis by PCR- SSCP

作　　者：李栋梁[1] 侯瑞生 王侃[1] 李辉[1] 杨红毅 高三友[1] 

机构地区：[1]河南省疾病预防控制中心,河南郑州450016 [2]河南出入境检验检疫局,河南郑州450003

出　　处：《中国卫生检验杂志》2014年第21期3127-3128,共2页Chinese Journal of Health Laboratory Technology

基　　金：河南省卫生厅医药卫生科研计划基金(201204116)

摘　　要：目的检测结核分枝杆菌耐药基因(Kat G,rpo B和rps L)突变,了解结核分枝杆菌对INH、RFP、SM的耐药状况。方法应用聚合酶链反应—单链构象多态性分析(PCR-SSCP)结合药物敏感试验,了解74例肺结核病人结核分枝杆菌基因突变情况和耐药情况。结果药敏实验耐药株52株(70.3%),其中耐INH41株(55.4%);耐RFP40株(54.1%);耐SM45株(60.1%)。同时耐3种药物的有36株(48.6%),耐2种药物的25株(33.8%),单耐药株13株(17.6%)。PCR-SSCP法对Kat G,rpo B和rps L基因突变的检出率分别为27.0%、51.4%、50.7%。耐INH、RFP和SM的kat G、rpo B和rps L基因突变的检出率分别为60.0%、89.7%和81.8%。结论 PCR-SSCP法检测结核分枝杆菌耐药基因突变是快速鉴定结核分枝杆菌耐药性的有效工具,与药敏试验联合应用可互补不足,提供更为详尽的结核分枝杆菌耐药情况,成为临床指导用药的有效工具。Objective To detect the drug - resistant genes （KatG, rpoB and rpsL） of mycobacterium tuberculosis. To understand the situation of drug - resistance of mycobacterium tuberculosis to INH, RFP and SM. Methods To investigate the condition of gene mutation and drug - resistance tests of M. tuberculosis clinical isolates from 74 patients with the tuberculosis by polymerase chain reaction - single stranded conformation polymorphism （PCR - SSCP） and drug sensitive test. Results There were 52 re- sisted strains（70.3% ）, 41 strains resisted INH（55.4% ）, 40 strains resisted RFP（54.1% ） and 45 resisted SM（60.1% ）, 36 strains resisted all the three drugs（33.8% ）, 25 strains resisted two drugs （33.8%） and 13 strains resisted single drug（ 17. 6% ）. The detection rates of mutation genes（KatG, rpoB and rpsL） were respectively 27.0%, 51.4% and 50.7%. That of the drug - resistant genes resisting SM, RFP, INH were 60.0%, 89.7% and 81.8%. Conclusion Detecting the resistance genes mutation of M. tuberculosis by PCR - SSCP was an efficient tool of identifying the drug resistance of M. Tuberculosis, and combined with drug sensitivity test, they can complement each other. Futhermore, it can also provide more detailed drug resistance condition of mycobacterium tuberculosis, and become an effective tool to guide clinical medication.

关 键 词：结核分枝杆菌 耐药性 基因突变 聚合酶链反应-单链构象多态性分析 

分 类 号：R446.5[医药卫生—诊断学]