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A sensitive loop‐mediated isothermal amplification (LAMP) method for detection of Renibacterium salmoninarum , causative agent of bacterial kidney disease in salmonids

S K Gahlawat; A E Ellis; B Collet
Journal of Fish Diseases · Vol. 32, Issue 6 · pp. 491-497 · 2009

Abstract

Loop‐mediated isothermal amplification (LAMP) is a novel technique for nucleic acid amplification with high specificity, sensitivity and rapidity and does not require expensive equipment or reagents. In the present study, we developed and evaluated a LAMP method for the rapid detection of Renibacterium salmoninarum causing the bacterial kidney disease in salmonids. This method was more sensitive than quantitative real‐time polymerase chain reaction (qPCR). Using DNA template extracted from cultured R. salmoninarum , the LAMP method gave an amplification signal from template diluted to 10 −8 while the limit of detection of qPCR was10 −7 . The LAMP method was also highly specific and did not amplify DNA purified from five other Gram‐positive and ‐negative bacterial fish pathogens. The method also worked well using extracts of macrophages infected with R. salmoninarum and kidney material from rainbow trout, which were positive for R. salmoninarum by qPCR and crude R. salmoninarum culture. There was some evidence for inhibitors of the LAMP reaction in the kidney samples, which was overcome by diluting the sample.

Bibliographic Information

JournalJournal of Fish Diseases
PublisherWiley
Publication Date2009-06-01
Publication Year2009
Volume32
Issue6
Pages491-497
Document TypeJournal Article
Print ISSN0140-7775
eISSN1365-2761
DOI10.1111/j.1365-2761.2009.01005.x
SubjectGeneral Aquaculture, Fisheries & Fish Science

Access Information

NARA Access Coverage1997-01-01~Current
Journal Homepagehttps://onlinelibrary.wiley.com/loi/13652761
Publisher PageOpen Publisher Page
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