环介导等温扩增技术在结核性胸膜炎快速诊断中的应用  

Application of loop-mediated isothermal amplification assay in the rapid diagnosis of patients with tuberculous pleuritis

作　　者：刘洋[1] 郭艳玲[2] 姜广路[3] 孙琦[4] 邢爱英[4] 张宗德[4] 

机构地区：[1]北京市结核病胸部肿瘤研究所,流行病学研究室,北京101149 [2]首都医科大学附属北京胸科医院检验科,北京101149 [3]首都医科大学附属北京胸科医院参比室,北京101149 [4]首都医科大学附属北京胸科医院分子生物学实验室,北京101149

出　　处：《临床肺科杂志》2015年第4期583-585,共3页Journal of Clinical Pulmonary Medicine

基　　金：北京市科技新星(No 2008A040);北京市优秀人才培养项目(No 2012D003034000006)

摘　　要：目的探讨针对hsp X基因设计引物的环介导等温扩增技术(LAMP)对结核性胸膜炎患者结核分枝杆菌的诊断价值。方法采用LAMP技术对结核分枝杆菌标准株H37Rv的DNA样品进行检测,评价LAMP技术的检测限;同时对8种非结核分枝杆菌菌株及阴沟肠杆菌和大肠埃希菌标准菌株进行鉴定,评价该方法的特异性。对58例结核性胸膜炎患者和32例肺癌患者胸腔积液样本结核分枝杆菌进行检测,评价LAMP技术的临床应用价值。结果 LAMP对倍比稀释的H37Rv的DNA样品检测结果为阳性,检测限为320a M。八种非结核分枝杆菌和2种普通菌菌株检测结果为阴性。LAMP对胸腔积液标本检测的灵敏度是75.8%(44/58),定量PCR为79.3%(46/58),无显著差别(χ2=0.25,P>0.05);均显著高于涂片法39.6%(23/58),差异具有统计学意义(χ2=17.39,P<0.01)。结论 LAMP技术是一种简单快速有效的等温扩增技术,由于其较高的灵敏度和特异度,可以对结核性胸膜炎患者胸腔积液标本快速鉴定。Objective To explore the application of loop-mediated isothermal amplification（ LAMP） assay in the rapid diagnosis of patients with tuberculous pleuritis. Methods Mycobacterium tuberculosis in pleural effusion from 58 patients with tuberculous pleuritis and 32 lung cancer patients were tested by LAMP. The specificity of this assay was evaluated using H37 Rv reference strains of mycobacterium tuberculosis,8 species of non-tuberculous mycobacteria（ NTM）,eterobacter cloacae,and E. coli. Results H37 Rv reference strain was successfully detected by this method,and there were no false-positive results of NTM,eterobacter cloacae and E. coli. The sensitivity of LAMP in detection of purified DNA from H37 Rv was 320 a M. The detection sensitivity for the pleural effusion from58 tuberculous pleuritis was 75. 8%（ 44 /58）,which was higher than smear microscopy（ 39. 6%）. There was no significant difference between the sensitivity of LAMP and that of quantitative real-time PCR（ χ^2= 0. 25,P〉0. 05）.Conclusion Because of its speed,simplicity,sensitivity and specificity,the TB hsp X LAMP assay is a potential gene diagnostic method for mycobacterium tuberculosis in patients with tuberculous pleuritis.

关 键 词：环介导等温扩增 结核分枝杆菌 胸腔积液 

分 类 号：R521.7[医药卫生—内科学]