Evaluation of a semi-nested PCR for detection of Renibacterium salmoninarum in samples from kidney, gill and ovarian fluid of Atlantic salmon broodfish

作者：Arnason Ivar Orn; Sigurdardottir Sunna; Kristmundsson Arni; Svansson Vilhjalmur; Gudmundsdottir Sigridur^*

来源：Icelandic Agricultural Sciences, 2013, 26: 49-57.

• Atlantic salmon
• bacterial kidney disease
• ELISA
• nested PCR
• Renibacterium salmoninarum
• semi-nested PCR

A semi-nested PCR (snPCR) for detection of Renibacterium salmoninarum that causes bacterial kidney disease (BKD) in salmonids was constructed. The efficacy of the snPCR was evaluated by comparison with nested PCR (nPCR) and two ELISA methods on kidney, ovarian fluid and gill samples collected from Atlantic salmon broodfish with escalating disease. The PCR methods using a conventional isolation kit identified equal numbers of positive samples, or 30%, with acceptable agreement. The ratio of positive kidney samples in PCR increased significantly, 42.5% in snPCR and 45% in nPCR, when an FTA minicard was used for DNA isolation. ELISA, using polyclonal antibodies, detected the highest number of positive samples (65%) and ELISA using monoclonal antibodies the lowest (17.5%). Ovarian fluid and gills gave inadequate results and cannot replace kidney samples for determination of R. salmoninarum infection in Atlantic salmon broodfish.

• 出版日期2013