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

• Main Authors: Ívar Örn Árnason 1982-, Sunna Sigurðardóttir 1949-, Árni Kristmundsson 1966-, Vilhjálmur Svansson 1960-, Sigríður Guðmundsdóttir 1950-
• Other Authors: Landbúnaðarháskóli Íslands
• Format: Article in Journal/Newspaper
• Language: English
• Published: 2013
• Subjects: Dýrasjúkdómar; Laxaætt; Atlantic salmon
• Online Access: http://hdl.handle.net/1946/19717

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. Semi-nested PCR (snPCR) aðferð var þróuð til greiningar á nýrnaveikibakteríunni Renibacterium salmoninarum. Greiningargeta snPCR prófsins í nýrna-, tálkna- og hrognavökvasýnum var metin út frá samanburði við nested PCR (nPCR) og tvær ELISA aðferðir í Atlantshafsklaklöxum með virka sýkingu. PCR aðferðirnar greindu sama hlutfall af jákvæðum sýnum með hefðbundinni DNA einangrunaraðferð, eða 30%, með góðu innbyrðis samræmi. Einangrun DNA á FTA pappír stuðlaði að greiningu marktækt fleiri jákvæðra sýna, 42.5% fyrir snPCR og 45% fyrir nPCR. Fjölstofna ELISA aðferð (pELISA) greindi flest jákvæð sýni (65%), en einstofna aðferðin (mELISA) fæst (17.5%). Tálkn og hrognavökvi henta ekki til skimunar á R. Salmoninarum í laxi.