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首页> 外文期刊>Applied Microbiology >Rapid Detection and Identification of Nontuberculous Mycobacterial Pathogens in Fish by Using High-Resolution Melting Analysis
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Rapid Detection and Identification of Nontuberculous Mycobacterial Pathogens in Fish by Using High-Resolution Melting Analysis

机译：高分辨率熔解法快速检测和鉴定鱼类中非结核分枝杆菌病原

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摘要

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 .
机译：鱼类中的分枝杆菌感染通常被称为鱼分枝杆菌病，与病原生物的具体身份无关。它们通常引起慢性疾病，有时可能导致高死亡率和严重的经济损失。据报道，近20种分枝杆菌感染鱼类。其中，通常认为海洋分枝杆菌，Fortuitum分枝杆菌和M. chelonae分枝杆菌是造成鱼分枝杆菌病的主要因素。由于没有快速，廉价的诊断测试，因此我们测试了高分辨率熔解分析（HRMA）的潜力，可以快速识别和区分与鱼类感染有关的几种分枝杆菌。通过分析16S-23S rRNA内部转录间隔区（ITS）的解链温度和解链谱，我们能够同时区分12种不同的物种。对纯化的海藻支原体和福特支原体DNA进行的敏感性测试表明，每个反应的检测限为10个基因组当量。在该程序中使用的引物未对16种对照非分枝杆菌属物种产生任何扩增信号，从而证明了它们对分枝杆菌属的特异性。

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