Wilson Teresa K, Douglas Marianne, Dunn Vivianne
Animal Health Laboratory, Department of Primary Industries Water and Environment, PO Box 46, Kings Meadows, Launceston, Tasmania 7249, Australia.
Dis Aquat Organ. 2019 Oct 30;136(3):219-226. doi: 10.3354/dao03407.
This work describes a new polymerase chain reaction (PCR) assay for rapid identification of the fish pathogens Tenacibaculum dicentrarchi, T. maritimum and T. soleae, 3 organisms which can cause tenacibaculosis in farmed salmonids. The selected primers amplified a 688 bp fragment for T. dicentrarchi, a 288 bp fragment of the T. maritimum and a 183 bp fragment of the T. soleae 16S rRNA genes. The PCR assay was shown to be both specific and sensitive with a detection limit of approximately 50 fg DNA for each species in the presence of competing DNA. The multiplex PCR allowed detection of each pathogen from pure or mixed cultures, where the different Tenacibaculum species can be difficult to distinguish phenotypically. Our results indicate that the specific primers and PCR method developed here provide sensitive and fast detection of T. dicentrarchi, T. maritimum and T. soleae alone or in combination.
这项研究描述了一种新的聚合酶链反应(PCR)检测方法,用于快速鉴定鱼类病原体——迪氏海杆菌、海生海杆菌和鳎海杆菌,这三种微生物可在养殖鲑科鱼类中引起海杆菌病。所选用的引物分别扩增出迪氏海杆菌688 bp的片段、海生海杆菌288 bp的片段以及鳎海杆菌16S rRNA基因183 bp的片段。在存在竞争性DNA的情况下,该PCR检测方法对每个物种的检测限约为50 fg DNA,显示出特异性和敏感性。多重PCR能够从纯培养物或混合培养物中检测出每种病原体,在这些培养物中,不同的海杆菌物种在表型上可能难以区分。我们的结果表明,此处开发的特异性引物和PCR方法能够单独或联合灵敏且快速地检测迪氏海杆菌、海生海杆菌和鳎海杆菌。