应用单链探针反向杂交试验检测结核分枝杆菌耐药性分析  

Rapid detection of mycobacterium tuberculosis resistance by PCR-LiPA

作　　者：刘靖元 李仁龙 徐亚军[2] 

机构地区：[1]深圳市龙岗区慢性病防治院,广东深圳518172 [2]深圳市龙岗区疾病预防控制中心,广东深圳518172

出　　处：《中国医药科学》2012年第13期26-28,共3页China Medicine And Pharmacy

摘　　要：目的探讨单链探针反向杂交试验检测结核分枝杆菌耐药性,评价此方法在耐药性检测中的临床应用价值。方法采用聚合酶链反应-单链探针反向杂交试验技术,PCR扩增50株结核分枝杆菌rpoB、katG、rpsL、rrs、embB耐药基因,将标记生物素的基因扩增产物与固定在硝酸纤维膜上的核酸探针杂交,通过链亲和素碱性磷酸酶,BICP/NBT显色系统检测分析基因突变,同时与传统药物敏感实验比对分析。结果 PCR-LiPA方法分析50株结核分枝杆菌临床分离株,耐RFP菌株中检出4株,耐INH菌株中检出5株,耐SM菌株中检出7株,SM敏感株检出1株,50株rrs基因均未检测到相应位点的突变;耐EMB菌株1株、EMB敏感株1株检测到embB基因突变。结论应用PCR-LiPA可快速、简便地检出部分结核分枝杆菌耐药株,可作为临床辅助诊断手段。Objective To detect mycobacterium tuberculosis resistance by LiPA,and to evaluate its clinical values. Methods 50 Mtb.were amplified the drug resistance gene ofrpoB, katG, rpsL, rrs, embB by PCR,and then hybridized with oligonucleotide probes on membrane,dyed by alkaline phosphatase and BICP/NBT system.At the same time,contrasted with the traditional drug sensitive test. Results 50 Mtb.isolate were performed the test from RFP-resistant Mtb.detected 4; of INH-resistant clinical isolate,5 were tested;of SM-resistant clinical isolate,7 were tested,1 Mtb.showed SM-resistant in all of SM-susceptible by LiPA;none Mtb.showed mutation in rrs genes;one EMB-resistant and one susceptible showed the mutation in embB genes. Conclusion PCR-LiPA might be a rapid,simple method for the detection of Mtb drug-resistance,it can be use for clinical detection as an assistant test.

关 键 词：结核分枝杆菌 耐药 异烟肼 利福平 乙胺丁醇 链霉素 PCR-LiPA ropB基因 KATG基因 RPSL基因 rrs基因 EMBB基因 

分 类 号：R-331[医药卫生]