Development and evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Flavobacterium psychrophilum.
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 x 10(1) to 2.0 x 10(9) copies/reaction within 70 min and revealed a good correlation between threshold times and gene copy number.
Fujiwara-Nagata
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E
Department of Fisheries, Kinki University, Nara, Japan. ns_ernag@nara.kindai.ac.jp
Eguchi
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M
D000818:Animals; D027101:DNA Topoisomerase IV / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005393:Fish Diseases / Q000175:diagnosis / Q000382:microbiology*; D045826:Flavobacteriaceae Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D005417:Flavobacterium / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics; D012487:Salmonidae; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D027101:DNA Topoisomerase IV