Comparative analysis of RAPD, RFLP and PFGE for the characterization of pathogenic vibrio parahaemolyticus isolated from seafood and coastal environment
Comparative analysis of RAPD, RFLP and PFGE for the characterization of pathogenic vibrio parahaemolyticus isolated from seafood and coastal environment
Date
2019
Authors
Minimol, V.A.
Kishore, P.
Nadella, R.K.
Panda, S.K.
Nayak, B.B.
Prasad, M.M.
Kumar, H.S.
Journal Title
Journal ISSN
Volume Title
Publisher
Society of Fisheries Technologists (India), Cochin
Abstract
Vibrio parahaemolyticus is ubiquitous in coastal marine
environment. Certain strains of V.
parahaemolyticus are pathogenic to humans and
harbors either or both thermostable direct hemolysin
(tdh) gene and tdh related hemolysin (trh) gene.
Pathogenic V. parahaemolyticus isolates from the
aquatic environment from Mumbai, India were
characterized with respect to their virulence genotypes
by employing random amplified polymorphic
DNA (RAPD), restriction fragment length polymorphism
(RFLP) and pulsed field gel electrophoresis
(PFGE). Out of 140 samples, twenty five samples
harboured virulent V. parahaemolyticus. Pandemic
group specific PCR assays viz., GSVP, GSOLD,
GSOK, ORF8 and PGS PCR showed differential
performances which revealed difficulties in the
confirmation of pandemic clone from environmental
samples. Results from the molecular characterization
employing RAPD and RFLP indicated that the PGS
PCR positive isolates (2) showed identical banding
pattern in RFLP analysis, whereas GS:OK positive
isolates (4) showed similar RAPD banding pattern
with 80% similarity among 11 pathogenic strains
studied. However, in whole genome comparison by
PFGE using notI restriction enzyme, these isolates
were scattered in different groups at 40% similarity
level. This study suggests that the genetically diverse
pathogenic strains of V. parahaemolyticus with characteristic
of pandemic clones are present in the
coastal environment.
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Citation
Fishery Technology 56(4):277-284