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A novel genotyping technique for distinguishing between Flavobacterium psychrophilum isolates virulent and avirulent to ayu, Plecoglossus altivelis altivelis (Temminck & Schlegel)
Author(s) -
FujiwaraNagata E,
Ikeda J,
Sugahara K,
Eguchi M
Publication year - 2012
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/j.1365-2761.2012.01368.x
Subject(s) - plecoglossus altivelis , biology , genotyping , virulence , polymerase chain reaction , genotype , microbiology and biotechnology , virology , pathogenicity , genetics , gene , fish <actinopterygii> , fishery
We developed a simple genotyping method for Flavobacterium psychrophilum for analysing two single nucleotide polymorphisms (SNPs) in the gyrA gene and to distinguish between isolates that are virulent and avirulent to ayu, Plecoglossus altivelis altivelis (Temminck & Schlegel). The genotyping method is an on/off switch assay and is based on the polymerase chain reaction technique with phosphorothioated primers. We classified 232 isolates from four families of fish (i.e. Plecoglossidae, Osmeridae, Cyprinidae and Salmonidae) into four genotypes (G‐C, A‐T, A‐C and G‐T). The G‐C type isolates exhibited strong pathogenicity to ayu, whereas the A‐T and G‐T types did not show any pathogenicity to this species. The A‐C type exhibited no or weak pathogenicity to ayu. These results indicate that genotyping F. psychrophilum isolates with two SNPs from gyrA can clearly distinguish between isolates potentially harmful to ayu (G‐C type) and those that are potentially not harmful or less harmful (A‐C, A‐T and G‐T type). The on/off switch assay provides a quick, simple, and very powerful DNA genotyping technique for F. psychrophilum isolates.