LAMP与RT-PCR法检测痰结核分枝杆菌的效果比较

Comparison of the Effect of LAMP and RT-PCR Assays for Rapid Detection of Mycobacterium Tuberculosis in Sputum Samples

目的比较环介导等温扩增技术(LAMP)和实时荧光PCR技术(RT-PCR)检测结核分枝杆菌的效果。方法可疑肺结核患者痰标本118份,分别采用痰涂片镜检、罗氏固体培养基培养、LAMP、RT-PCR方法进行结核分枝杆菌检查。结果结核分枝杆菌阳性检出率痰涂片镜检38%、罗氏固体培养基培养41%、LAMP45%、RT-PCR 55%,4种方法阳性检出率比较差异有统计学意义(P<0.05);以常规罗氏固体培养法为标准,LAMP法的灵敏性和特异性分别为92.0%(46/50)和86.8%(59/68),与罗氏培养法比较差异无统计学意义(P>0.05),一致性强(Kappa=0.777);PCR法的灵敏性和特异性分别为86.0%(43/50)和66.2%(45/68),与罗氏培养法比较差异有统计学意义(P<0.01)。结论 RT-PCR检测法阳性率最高,但特异性低;LAMP法灵敏性和特异性较高,与传统培养法一致性好,操作简便易行、快速准确。

To compare the effect of loopmediated isothermal amplification （LAMP）and real-time reverse transcription polymerase chain reaction （RTPCR）assay for the detection of mycobacterium tuberculosis in sputum samples. Methods One hundred and eighteen sputum samples from suspicious tuberculosis （riB）patients weredetected by the four methods including smear microscopy, LJ solid culture, LAMP and RT-PCR assay. Results The positive detection rates of smear microscopy, LJ culture, LAMP and RT-PCR were 38% ,41% ,45% and 55%, respectively. There were significant differences in the positive rate among the four methods （ P 〈 0.05 ）. Compared to the L-J solid culture, the sensitivity and specificity of LAMP were 92.0% （46/50）and 86.8% （59/68）, and there was no significant difference between the two methods （ P 〉 0.05 ）, but with high concordance （ Kappa = 0. 777 ）. The sensitivity and specificity of RTPCR assay were 86.0% （43/50） and 66.2% （45/68） ,and showed significant differences between the LJ solid culture and RTPCR assay（P 〈0. 01 ）. Conclusion RTPCR assay shows the highest positive rate in the four methods,but of lowest specificity. LAMP may provide a simple,rapid,sensitive and cost-effective method for the detection of pulmonary TB,with high sensitivity and specificity,which has a high concordance with traditional culture.