环介导等温扩增法快速检测结核分枝杆菌的临床应用评估

中国防痨杂志 ›› 2012, Vol. 34 ›› Issue (7): 413-418.

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环介导等温扩增法快速检测结核分枝杆菌的临床应用评估

陈涛周琳周杰李海成江勇彭东东周志刚彭建明文力何超文钱明孙毅凡石磊钟球

106130 广州，广东省结核病控制中心参比实验室（陈涛、周琳、李海成、江勇、钱明、钟球）；广东省佛山市第四人民医院（周杰）；广东省汕头市结核病防治所（彭东东）；广东省东莞市慢性病防治院（周志刚）；广东省惠州市结核病防治所（彭建明）；广东省江门市结核病防治所（文力）；广州市番禺区慢性病防治站（何超文）；暨南大学医学院微生物与免疫学教研室（孙毅凡）；华南理工大学轻工与食品学院（石磊）

收稿日期:2012-05-16 出版日期:2012-07-10 发布日期:2012-07-06
通信作者: 钟球；周琳 E-mail: zhongqiu@vip.163.com；gdtb_bg@vip.163.com
基金资助:
“十二五”国家科技重大专项（2012ZX10004903）;“十二五”广东省“结核病防治转化”医学重点实验室

Clinical application of loop-mediated isothermal amplification method in rapid detection of Mycobacterium tuberculosis

CHEN Tao, ZHOU Lin, ZHOU Jie, LI Hai-cheng, JIANG Yong, PENG Dong-dong, ZHOU Zhi-gang, PENG Jian-ming, WEN Li, HE Chao-wen, QIAN Ming, SUN Yi-fan, SHI Lei, ZHONG Qiu

Center for Tuberculosis Control and Prevetion of Guangdong Province，Guangzhou 106130,China

Received:2012-05-16 Online:2012-07-10 Published:2012-07-06
Contact: ZHONG Qiu；ZHOU Lin E-mail: zhongqiu@vip.163.com；gdtb_bg@vip.163.com

摘要/Abstract

摘要： 目的进行环介导等温扩增法（LAMP法）快速检测结核分枝杆菌的临床验证与应用，评估该方法在结核病临床诊断中的效能。方法选取广东6地市专业结核病防治机构（简称“结防机构”）的肺部不同疾病及健康人员的痰标本2129份，分别采用直接涂片法、罗氏固体培养法及LAMP法检测痰标本中结核分枝杆菌；另广州、汕头两市337例结核病患者的痰标本浓缩处理后进行实时荧光定量（qPCR）及LAMP两种方法检测。结果 2129例痰标本中，直接涂片法、罗氏固体培养法和LAMP法的阳性检出率分别为13.2%（282/2129）、21.7%（463/2129）和20.3%(432/2129)；同罗氏培养法相比，LAMP法灵敏度、特异度、阳性和阴性预测值分别为89.1%(432/485)、95.5%(1570/1644)、85.4%(432/506)和96.7%(1570/1623)；337例浓缩处理后的痰标本qPCR阳性率为35.0% (118/337)，LAMP阳性率为35.9% (121/337)；LAMP法同直接涂片法检测、罗氏固体培养法和qPCR法比较，检测结果实际一致率分别为87.9%（Kappa值＝0.612）、96.1%（Kappa值＝0.89）和91.4%（Kappa值＝0.812）。结论 LAMP法用于痰标本中结核分枝杆菌检测，其效能同罗氏培养法及qPCR法相当，同直接涂片法相比，LAMP法能够大幅提高阳性检出率，具有很好的临床应用和推广前景。

关键词: 分枝杆菌,结核, 核酸扩增技术

Abstract: Objective To evaluate the diagnostic value of a loop-mediated isothermal amplification (LAMP) method for rapid detection of Mycobacterium tuberculosis. Methods 2129 sputum samples of suspected TB patients from 6 cities in Guangdong province and 337 enriched sputum samples randomly selected from Guangzhou and Shantou cities were collected. M. tuberculosis in 2129 sputum samples were detected by smear, Roche solid culture and LAMP methods. M. tuberculosis in 337 sputum samples were detected by quantitative polymerase chain reaction (qPCR) and LAMP methods. Results For 2129 sputum samples, the positive rates of smear, culture and LAMP were 13.2%（282/2129）, 21.7%（463/2129）and 20.3%(432/2129), respectively. Compared with Roche culture method, sensitivity, specificity, positive predictive value and negative predictive value of the LAMP method were 89.1%(432/485), 95.5%(1570/1644), 85.4%(432/506) and 96.7%(1570/1623), respectively. For 337 enriched sputum samples, the positive rates of qPCR and LAMP were 35.0% (118/337) and 35.9% (121/337), respectively. Compared LAMP with smear, culture and qPCR, the consistent rate of the results were 87.9%（K=0.612）, 96.1%（K=0.89）and 91.4%（K=0.812）, respectively. Conclusion The LAMP method was equivalent to culture and qPCR, superior to smear method in detecting M. tuberculosis of sputum specimens. In addition, LAMP method showed the advantages of rapid, simple operation and no equipment, indicating good prospects for clinical application and promotion in the diagnosis of tuberculosis.

Key words: Mycobacterium tuberculosis, Nucleic acid amplification techiques

陈涛周琳周杰李海成江勇彭东东周志刚彭建明文力何超文钱明孙毅凡石磊钟球. 环介导等温扩增法快速检测结核分枝杆菌的临床应用评估[J]. 中国防痨杂志, 2012, 34(7): 413-418.

CHEN Tao, ZHOU Lin, ZHOU Jie, LI Hai-cheng, JIANG Yong, PENG Dong-dong, ZHOU Zhi-gang, PENG Jian-ming, WEN Li, HE Chao-wen, QIAN Ming, SUN Yi-fan, SHI Lei, ZHONG Qiu. Clinical application of loop-mediated isothermal amplification method in rapid detection of Mycobacterium tuberculosis[J]. Chinese Journal of Antituberculosis, 2012, 34(7): 413-418.

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环介导等温扩增法快速检测结核分枝杆菌的临床应用评估

Clinical application of loop-mediated isothermal amplification method in rapid detection of Mycobacterium tuberculosis

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