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首页> 外文期刊>Fisheries and Aquatic Sciences >Species-Specific Duplex PCR for Detecting the Important Fish Pathogens Vibrio anguillarum and Edwardsiella tarda
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Species-Specific Duplex PCR for Detecting the Important Fish Pathogens Vibrio anguillarum and Edwardsiella tarda

机译：用于检测重要鱼类病原弧菌和塔氏爱德华氏菌的特定物种双链PCR

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摘要

Vibriosis caused by Vibrio anguillarum and edwardsiellosis caused by Edwardsiella tarda are septicemic diseases of many commercially important freshwater and marine fishes, and threaten the aquaculture industry in Korea. Early diagnosis and accurate identification of these two bacterial species could help to prevent these diseases and minimize the damage to cultured marine species. This study designed a duplex polymerase chain reaction (PCR) method for the simultaneous detection of two major fish pathogens: V. anguillarum and E. tarda. Each pair of oligonucleotide primers exclusively amplified the target groEL gene of the specific microorganism. Twenty-two Vibrio and ten non-Vibrio enteric species were used to check the specificity of the primers, which were found to be highly specific for the target species, even among closely related species. The detection limit was 400 pg for V. anguillarum and 4 ng for E. tarda when mixed purified DNA was used as the template. This assay showed high specificity and sensitivity in the simultaneous detection of V. anguillarum and E. tarda from artificially inoculated seawater and fish.
机译：鳗弧菌引起的弧菌病和塔氏爱德华氏菌引起的埃德氏病是许多商业上重要的淡水和海水鱼类的败血病，并威胁着韩国的水产养殖业。对这两种细菌的​​早期诊断和准确识别有助于预防这些疾病，并最大程度地减少对养殖海洋物种的损害。这项研究设计了一种双工聚合酶链反应（PCR）方法，用于同时检测两种主要鱼类病原体：鳗弧菌和塔氏大肠杆菌。每对寡核苷酸引物专门扩增特定微生物的靶标groEL基因。使用22种弧菌和10种非弧菌肠溶菌种来检查引物的特异性，发现这些引物对目标菌种具有高度特异性，即使在密切相关的菌种中也是如此。当使用混合的纯化DNA作为模板时，对鳗弧菌的检测限为400 pg，对E鳗的检测限为4 ng。该检测方法在从人工接种的海水和鱼类中同时检测鳗弧菌和塔氏大肠杆菌时显示出很高的特异性和敏感性。

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