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目的建立结核分枝杆菌实时荧光核酸恒温扩增检测方法(SAT-TB),并探讨其在结核病诊断中的应用。方法设计特异性引物和探针,构建SAT检测体系,并优化。通过不同浓度H37Rv标准菌株检测评估其灵敏性。结果结核分枝杆菌SAT检测灵敏度为10CFU/ml;非结核分枝杆菌及常规细菌未检出TB RNA。肺结核患者痰液标本中结核分枝杆菌SAT检出率为54%,显著高于痰培养法的40%(P<0.05)及痰涂片法的34%(P<0.01)。结论 SAT方法应用于结核分枝杆菌检测,快速方便、灵敏度高、特异性强,可以应用于临床结核病诊断。
Objective To establish a real-time fluorescent nucleic acid thermostable amplification of Mycobacterium tuberculosis detection method (SAT-TB) and to explore its application in the diagnosis of tuberculosis. Methods Specific primers and probes were designed to construct SAT detection system and optimized. Sensitivity was assessed by testing H37Rv standard strains at various concentrations. Results The detection sensitivity of Mycobacterium tuberculosis SAT was 10 CFU / ml. TB RNA was not detected in Mycobacterium tuberculosis and routine bacteria. The detection rate of Mycobacterium tuberculosis SAT was 54% in sputum samples of patients with pulmonary tuberculosis, which was significantly higher than 40% (P <0.05) of sputum culture method and 34% (P <0.01) of sputum smear method. Conclusion The SAT method is applied to the detection of Mycobacterium tuberculosis, which is rapid, convenient, sensitive and specific. It can be used in clinical diagnosis of tuberculosis.