结核分支杆菌耐利福平耐药基因检测研究  

Defection rifampin-resistant Mycobacterium tuberculosis

作　　者：张晓娟[1] 张小刚[2] 董亚俊 何秀云[2] 吕树峰 陈小冬 

机构地区：[1]沈阳市胸科医院检验科,辽宁沈阳110044 [2]解放军309医院结核病研究室 [3]辽宁省朝阳市结核病医院

出　　处：《中国现代医学杂志》2004年第3期86-88,共3页China Journal of Modern Medicine

摘　　要：目的　评价应用DNA序列分析方法和聚合酶链反应 -单链构象多态性 (PCR -SSCP)技术检测rPOB基因突变在结核分支杆菌耐利福平 (RFP)耐药性测定中的应用价值。方法　采用DNA序列分析法和PCR -SSCP法对 80株结核分支杆菌临床分离株 (其中药物敏感株 32株 ,耐RFP或含耐RFP耐多药株 4 8株 )rpoB基因核心区域的突变情况进行检测。 结果　以结核分支杆菌H37RV为对照 ,所有 32株药物敏感株的rPOB基因均无突变。用DNA序列分析方法检测 4 8株耐药株中 4 4株发生突变 ,敏感性为 91.7% (44 /48) ,其中最常见的突变位点是 5 31位丝氨酸和 5 2 6位组氨酸。PCR -SSCP检测 4 8株耐RFP分离株中 ,4 5株rPOB基因SSCP图谱异常 ,其敏感性为 93.1% (45 /48)。结论　DNA序列分析可指导临床用药 ,PCR -SSCP可快速检测结核分支杆菌RFP耐药性。Objective: Detection rPOB geng mutations in refampin-resistant Mycobacterium tuberculosis using Polymerase Chain reaction-single-strand conformation polymorphi RFP(PCR-SSCP) and DNA sequencing. Methods:80 clinical isolates of Mycobacterium tuberculosis (32 drug susceptible,48 rifampin-resistance or multidrug resistance including rifampin) by DNA sequencing and PCR-SSCE.Results: No mutation was found in 32 rif-rensitive strains by DNA sequencing. While 44 of 48 rif-rensitive strains had mutations rifampin resistant determination region of rpoB gene, the mutation rate was 91.7%(44/48),531-Ser and 526-His were the most common positions to be substituted.32 rif-rensitive strains had same PCR-SSCP gelprofile with Mycobacterium tuberculosis H37Rv as control, while 45 of 48 rif-rensitive strains showed different SSCP. The specificity is 93.1%. Conclusion:DNA sequencing is significant in drug choosing for tuberculosis treatment. Rifampin-resistant of Mycobacterium tuberculosis is rapidly and accurately detected using PCR-SSCP.

关 键 词：DNA序列分析 PCR-SSCPrpoB基因 结核分支杆菌 

分 类 号：Q523.8[生物学—生物化学]