rDNA探针杂交检测病人痰标本中结核杆菌的研究  

The study of detecting sputum specimen of patients of pulmonary tuberculosis by rDNA probe

作　　者：张灵霞[1] 庄玉辉[1] 杨华卫[1] 李邦印[1] 夏湘萱[1] 

机构地区：[1]解放军309医院结核病研究室,北京100091

出　　处：《中国防痨杂志》2000年第3期127-129,共3页Chinese Journal of Antituberculosis

摘　　要：应用结核分枝杆菌特异的rDNA探针杂交检测痰标本中的结核杆菌rDNA ,评价其在临床标本检测中的应用价值。方法　用引物b对结核分枝杆菌 1 6S - 2 3SrDNA间隔区序列进行扩增 ,同时加入生物素标记 ,制成 2 50bp的rDNA探针。对该探针的敏感性、特异性进行了研究 ,并对 90份结核病人 ,30份非结核病人痰标本的PCR产物进行了斑点杂交检测。结果　rDNA探针检测引物bPCR扩增产物的敏感性为 1 0 0pg。rDNA探针与受试 2 4种分枝杆菌和 1 1种非分枝杆菌引物bPCR扩增产物杂交只有结核分枝杆菌、胃分枝杆菌为阳性杂交 ,特异性较高。而rDNA探针对 90份结核病人痰标本引物b扩增产物检测的阳性率为 80 .2 % ,高于PCR扩增产物电泳检测结果 (64% )。rDNA探针与 30份非结核病人痰标本杂交结果均为阴性杂交。结论　rDNA探针与引物bPCR扩增相结合能提高结核病人痰标本检测的敏感性与特异性。Objective Using rDNA probe detecting mycobacteria to establish a new method detecting clinical specimen.Methods Using special primer b to produce PCR probe labeled with biotin.Specificity and sensitivity of the probe detecting primer b PCR products were studied,the positive rate of this probe detecting clinical specimen were also studied.Results The sensitivity was 100pg/μl,only M.tuberculosis and M.gastri were positive hybridization.The probe hybridized with prime b PCR products of 90 sputum specimen of pulmonary tuberculosis patients,positive rate was 80.2%.Conclusion rDNA probe with PCR was a practical methods in detecting M.tuberculosis.

关 键 词：rDNA探针 结核分枝杆菌 聚合酶链反应 肺结核 

分 类 号：R446.5[医药卫生—诊断学]