Development of an in-house loop-mediated isothermal amplification (LAMP) assay for detection of Mycobacterium tuberculosis and evaluation in sputum samples of Nepalese patients
Development of an in-house loop-mediated isothermal amplification (LAMP) assay for detection of Mycobacterium tuberculosis and evaluation in sputum samples of Nepalese patients
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DOI：
10.1099/jmm.0.47499-0
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发表时间：
2008-04-01
影响因子：
3
通讯作者：
Suzuki, Yasuhiko
中科院分区：
文献类型：
--
作者：
Pandey, Basu Dev;Poudel, Ajay;Suzuki, Yasuhiko
许多核酸扩增试验(NAAS)已用于检测临床标本中的结核杆菌，用于结核病的诊断。其中，环介导的等温扩增(LAMP)是一种具有优越等温反应特性的NAA。本研究选择了一组针对结核分枝杆菌16S rRNA基因的高灵敏度的特异性引物，建立了一套LAMP系统(MTB-LAMP)。使用该系统，总共对200份尼泊尔患者的痰样本进行了调查。培养阳性标本MTB-LAMP的敏感性为100%(96/96)，培养阴性标本的特异性为94.2%(98/104，95%可信区间90.5~97.9%)。MTB-LAMP的阳性预测值为94.1%，阴性预测值为100%。这些结果表明，这种MTB-LAMP方法可能被证明是一种强有力的结核病早期诊断工具。
A number of nucleic acid amplification assays (NAAs) have been employed to detect tubercle bacilli in clinical specimens for tuberculosis (TB) diagnosis. Among these, loop-mediated isothermal amplification (LAMP) is an NAA possessing superior isothermal reaction characteristics. In the present study, a set of six specific primers targeting the Mycobacterium tuberculosis 16S rRNA gene with high sensitivity was selected and a LAMP system (MTB-LAMP) was developed. Using this system, a total of 200 sputum samples from Nepalese patients were investigated. The sensitivity of MTB-LAMP in culture-positive samples was 100 % (96/96), and the specificity in culture-negative samples was 94.2 % (98/104, 95 % confidence interval 90.5-97.9 %). The positive and negative predictive values of MTB-LAMP were 94.1 and 100 % respectively. These results indicate that this MTB-LAMP method may prove to be a powerful tool for the early diagnosis of TB.