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目的对目前临床上用于检测结核分枝杆菌(MTB)及其耐药性的传统及分子生物学方法进行系统评价,为设计合理的诊断路径提供指导意义。方法对标准菌株(H37Rv)、敏感、单耐及耐多药MTB进行梯度浓度稀释后,作为评估的实验样本,从检测结果的准确性、检测周期、最低检出限方面对不同方法进行评价。结果MTB检测方面,XpertMTB/RIF检测MTB的检出下限(3×10~2cfu)与培养法相当,比基因芯片和GenoType~ MTBDRplus的检出下限低;耐药性检测方面,在MTB含量不足(≤3×10~2cfu)的情况下XpertMTB/RIF对利福平(RFP)的检测存在假阳性结果,GenoType~ MTBDRplus检测3×10~4cfu链霉素耐药MTB时出现RFP和异烟肼(INH)耐药的假阳性结果;检测周期方面,XpertMTB/RIF在2 h内可完成报告,基因芯片和GenoType~ MTBDRplus在当天8 h内可完成报告,培养及比例法药敏需4~8周可报告结果。结论目前不同的MTB及耐药检测方法各有优缺点,在临床应用方面应综合考虑患者的病情、经济状况等选取合理的诊断路径。
OBJECTIVE: To systematically evaluate the current clinical and molecular methods for detection of Mycobacterium tuberculosis (MTB) and its drug resistance, and to provide guidance for the design of rational diagnostic pathways. Methods Different dilutions of standard strains (H37Rv), sensitive, single-resistant and multi-drug resistant MTB were used as experimental samples to assess the accuracy of test results, test cycle and minimum detectable limit. Results MTB detection, XpertMTB / RIF detection of MTB detection limit (3 × 10 ~ 2cfu) and culture method, gene chip and GenoType ~ MTBDRplus lower detection limit; drug resistance testing, MTB content is low (≤3 × 10 ~ 2cfu), there was a false positive result for the detection of rifampicin (RFP) by XpertMTB / RIF. When GenoType ~ MTBDRplus detected 3 × 10 ~ 4cfu streptomycin-resistant MTB, Hydrazine (INH) resistance test results; XpertMTB / RIF in the detection cycle can be completed within 2 h report, gene chip and GenoType ~ MTBDRplus within 8 h on the report can be completed, the culture and the proportion of drug sensitive need 4 ~ 8 weeks to report the results. Conclusion At present, different MTB and drug resistance test methods have their own advantages and disadvantages. In clinical application, the reasonable diagnostic path should be selected considering the patient’s condition and economic status.