【摘要】 目的　研制线性探针分析法检测结核分枝杆菌 (结核菌 )rpoB基因耐药突变试剂盒。方法　设计 15条结核菌rpoB基因野生型和突变型的线性探针 ,固定尼龙膜上 ;在rpoB基因引物上标记生物素 ,扩增结核菌DNA ,获得生物素标记扩增产物 ,与rpoB基因线性探针杂交 ,加入标记过氧化物酶的亲合素 ,再加四甲基联苯胺显色。结果　应用线性探针分析法检测 87株结核菌 ,其与rpoB基因直接测序法、传统药敏法符合率分别为 98 8% (82 / 83)和 94 3% (82 / 87)。结论　线性探针分析法检测rpoB耐药基因突变是一种操作简便、特异性高、敏感性高 ,易开展的方法。更多还原

【Abstract】 Objective To make a line probe assay (LiPA) kits for detecting the mutations of rpoB rifampin resistance relevant gene in Mycobacterium tuberculosis Methods fifteen single strand specific probes designed to detect mutated and/or wild rpoB gene in Mycobacterium tuberculosis were spotted and fixed on nylon membranes PCR products labeled with biotin were obtained by using up stream primer labeled with biotin, probes hybridized with PCR products and analyzed with POD and TMB The results were concluded through the naked eye directly Results 87 strains Mycobacterium tuberculosis were detected by Line probe assay kits.Hybridizing results conparing with sequencing results and traditional susceptibility test were 98 8%(82/83),94 3%(82/87) respectively Conclusion Line probe assay kit was a good kits for rapid detecting rifampin resistant tuberculosis. It was suited to develop in clinical laboratory更多还原