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Development and evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Flavobacterium psychrophilum
Authors:E Fujiwara-Nagata  M Eguchi
Institution:Department of Fisheries, Kinki University, Nara, Japan
Abstract:Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease and rainbow trout fry syndrome of salmonids. The pathogen has been reported from all regions in the world involved in salmonid aquaculture, but also from natural fresh-water environments. We established a quantitative loop-mediated isothermal amplification of DNA (LAMP) method to estimate quantities of F. psychrophilum . LAMP primers were designed based on the sequence of the DNA topoisomerase IV subunit B gene, parE , of F. psychrophilum. parE LAMP exhibited a high specificity for the parE gene of F. psychrophilum but not for other related species. parE LAMP detected the gene in a wide range of concentrations from 2.0 × 101 to 2.0 × 109 copies/reaction within 70 min and revealed a good correlation between threshold times and gene copy number.
Keywords:bacterial cold-water disease              Flavobacterium psychrophilum            loop-mediated isothermal amplification of DNA  quantification
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