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基于环介导等温扩增技术的实验室检测方法在快速鉴定固体培养基上生长的结核分枝杆菌复合群中的应用。

Applicability of in-house loop-mediated isothermal amplification for rapid identification of Mycobacterium tuberculosis complex grown on solid media.

机构信息

National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand.

出版信息

Jpn J Infect Dis. 2013;66(3):249-51. doi: 10.7883/yoken.66.249.

Abstract

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.

摘要

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。

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