National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand.
Jpn J Infect Dis. 2013;66(3):249-51. doi: 10.7883/yoken.66.249.
A simple, rapid, and low-cost identification method is required in tuberculosis high-burden countries. We report the applicability of in-house loop-mediated isothermal amplification (LAMP) targeting 16S ribosomal RNA for the rapid identification of Mycobacterium tuberculosis complex grown on Lowenstein-Jensen media. Eighty acid-fast staining-positive clinical isolates were selected and used to evaluate the LAMP assay in comparison with polymerase chain reaction and conventional culture-based tests. The LAMP assay identified 60 M. tuberculosis isolates from 80 clinical isolates using simple heat-extracted DNA directly from the colony suspension. The results were in complete agreement with those obtained using the other methods, and the utility of the direct LAMP assay from a colony was demonstrated. The LAMP assay appears to be a practical and low-cost method that can be used for the rapid identification of M. tuberculosis isolates and suitable for endemic low-resource settings.
在结核病负担重的国家,需要一种简单、快速且低成本的鉴定方法。我们报告了针对 16S 核糖体 RNA 的内部环介导等温扩增 (LAMP) 在快速鉴定在 Lowenstein-Jensen 培养基上生长的结核分枝杆菌复合体方面的适用性。选择了 80 株抗酸染色阳性的临床分离株,并用其评估 LAMP 检测与聚合酶链反应和传统基于培养的检测方法的比较。LAMP 检测使用简单的热提取 DNA 直接从菌落在悬浮液中鉴定了 80 株临床分离株中的 60 株结核分枝杆菌分离株。结果与其他方法完全一致,并且证明了直接从菌落进行 LAMP 检测的实用性。LAMP 检测似乎是一种实用且低成本的方法,可用于快速鉴定结核分枝杆菌分离株,适用于流行地区的资源匮乏环境。