Phung Thu Nguyet, Caruso Domenico, Godreuil Sylvain, Keck Nicolas, Vallaeys Tatiana, Avarre Jean-Christophe
Institute of Biotechnology, Vietnam Academy of Science and Technology, Cau Giay, Hanoi, Vietnam.
Appl Environ Microbiol. 2013 Dec;79(24):7837-45. doi: 10.1128/AEM.00822-13. Epub 2013 Oct 11.
Mycobacterial infections in fish are commonly referred to as piscine mycobacteriosis, irrespectively of the specific identity of the causal organism. They usually cause a chronic disease and sometimes may result in high mortalities and severe economic losses. Nearly 20 species of Mycobacterium have been reported to infect fish. Among them, Mycobacterium marinum, M. fortuitum, and M. chelonae are generally considered the major agents responsible for fish mycobacteriosis. As no quick and inexpensive diagnostic test exists, we tested the potential of high-resolution melting analysis (HRMA) to rapidly identify and differentiate several Mycobacterium species involved in fish infections. By analyzing both the melting temperature and melting profile of the 16S-23S rRNA internal transcribed spacer (ITS), we were able to discriminate 12 different species simultaneously. Sensitivity tests conducted on purified M. marinum and M. fortuitum DNA revealed a limit of detection of 10 genome equivalents per reaction. The primers used in this procedure did not lead to any amplification signal with 16 control non-Mycobacterium species, thereby demonstrating their specificity for the genus Mycobacterium.
鱼类中的分枝杆菌感染通常被称为鱼类分枝杆菌病，无论致病生物体的具体种类如何。它们通常会引发一种慢性疾病，有时可能导致高死亡率和严重的经济损失。据报道，近20种分枝杆菌可感染鱼类。其中，海分枝杆菌、偶然分枝杆菌和龟分枝杆菌通常被认为是导致鱼类分枝杆菌病的主要病原体。由于不存在快速且廉价的诊断测试方法，我们测试了高分辨率熔解分析（HRMA）在快速鉴定和区分几种与鱼类感染有关的分枝杆菌物种方面的潜力。通过分析16S - 23S rRNA内部转录间隔区（ITS）的熔解温度和熔解曲线，我们能够同时鉴别出12种不同的物种。对纯化的海分枝杆菌和偶然分枝杆菌DNA进行的敏感性测试显示，每个反应的检测限为10个基因组当量。本实验所使用的引物对16种对照非分枝杆菌物种未产生任何扩增信号，从而证明了它们对分枝杆菌属的特异性。
