Genotyping of Flavobacterium psychrophilum using PCR-RFLP analysis

Genetic variability among 242 strains of Flavobacterium psychrophilum was characterized using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) analysis. Universal Primers GYR-1 and GYR-1R, which were designed to amplify the gyrase subunit B gene (gyrB), yie...

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Bibliographic Details
Published in:Diseases of aquatic organisms 2003-10, Vol.56 (3), p.207-214
Main Authors: IZUMI, Shotaro, ARANISHI, Futoshi, WAKABAYASHI, Hisatsugu
Format: Article
Language:English
Subjects:
Animals
Bacteriology
Base Sequence
Biological and medical sciences
DNA Primers
Electrophoresis, Agar Gel
Fishes - microbiology
Flavobacterium - genetics
Flavobacterium psychrophilum
Fundamental and applied biological sciences. Psychology
Genetic Variation
Geography
Japan
Microbiology
Molecular Sequence Data
Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
Sequence Analysis, DNA
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Summary:Genetic variability among 242 strains of Flavobacterium psychrophilum was characterized using polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) analysis. Universal Primers GYR-1 and GYR-1R, which were designed to amplify the gyrase subunit B gene (gyrB), yielded a 1178 bp PCR product encoding gyrB and a 290 bp PCR product of anonymous DNA from all F. psychrophilum strains tested. In the RFLP analysis of the anonymous 290 bp DNA marker, the restriction enzyme HinfI generated 2 cleavage patterns (Genotypes A and B). Genotype A was found only in isolates from ayu (n = 109), while Genotype B was found in isolates from coho salmon (n = 11), ayu (n = 35), rainbow trout (n = 43) and other fishes (n = 44). In the second experiment, Primers PSY-G1F and PSY-G1R specific for F. psychrophilum, were used to amplify gyrB. The specific primer pair amplified the expected size (1017 bp) PCR product from all F. psychrophilum strains. In the RFLP analysis of the gyrB, the restriction enzyme RsaI produced 2 genotypes, R and S. Genotype R was found in isolates from coho salmon (n = 6), ayu (n = 27), rainbow trout (n = 39) and other fishes (n = 4). Genotype S was found in isolates from coho salmon (n = 5), ayu (n = 117), rainbow trout (n = 4) and other fishes (n = 40).
ISSN:0177-5103
1616-1580
DOI:10.3354/dao056207