环介导等温扩增技术快速检测痰标本中结核分枝杆菌的初步评价

目的评价环介导等温扩增技术（loop-mediated isothermal amplification,LAMP）技术在结核分枝杆菌诊断中的应用价值。方法收集246份疑似肺结核患者的痰标本,分别进行涂片镜检,罗氏培养以及实时定量PCR和LAMP法检测。结果涂片镜检,罗氏培养实时定量PCR和LAMP法对结核分枝杆菌的检出率分别为30.89%,45.53%,54.47%,57.32%。以罗氏培养结果作为金标准,LAMP法诊断的敏感性为96.19%（101/105）,特异性为76.87%（103/134）,两种方法检测结果的一致性是Κ=0.71,两法具有很好的吻合性。以实时定量法为标准,LAMP法的敏感性为90.07%（127/141）,特异性为93.33%（98/105）,两法检测结果的一致性是κ=0.83,P〈0.001。结论 LAMP法检测结核分枝杆菌具有较高的敏感性和特异性,与罗氏培养和RT-PCR有着较高的吻合性,很适合作为临床检测项目开展。

Evaluation of Loop-mediated isothermal amplification for the rapid detection of M. tuberculosis in sputa

Objective To evaluate the diagnostic value of loop-mediated isothermal amplification（LAMP）in the diag nosis of Mycobacterium tuberculosis. Methods Collected 246 sputa from suspected tuberculosis patients and conducted smear microscopy,IJ culture,real-time quantitative PCR （RT-PCR）and LAMP assay. Results The detection rate of smear microscopy, LJ culture, RT-PCR and LAMP assay for Mycobacterium tuberculosis were 30. 89%, 45. 53%, 54.47% and 57.32%, respectively. Comparing with culture method, the sensitivity and the specificity of the LAMP assay was 96. 19% （101/105） and 76. 87% （103/134）, respectively. The two assays were in good consistency （K= 0.71）. Comparing with RT-PCR,the sensitivity and specificity of the LAMP was 90.07% （127/141） and 93.33%（98/ 105）, respectively. The two assays were in good consistency（K=0.83） . Conclusion LAMP assay for rapid detection of Mycobacterium tuberculosis with high sensitivity and specificity,has high consistency with culture and RT-PCR,and is very suitable for routine clinical test.