rDNA探针杂交检测病人痰标本中结核杆菌的研究

目的 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16S-23SrDNA间隔区序列进行扩增，同时加入生物素标记，制成250bp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100pg。rDNA探针与受试24种分枝杆菌和11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。

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

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.