STUDIES ON THE INCIDENCE OF VIBRIO VULNIFICUS IN AQUATIC FOODS AND ENVIRONMENTAL SAMPLES
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Date
2016-05-16
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PVNR TVU
Abstract
The present study was undertaken to standardize PCR assay for detection of Vibrio vulnificus from aquatic foods and environmental samples and compare with conventional cultural methods. Primers derived from vvhA and gyrB genes gave specific amplification at 519 bp and 285 bp for Vibrio vulnificus respectively.
Four different template preparation methods viz. genomic DNA extraction, heat lysis, lysis buffers-1 and 2 were compared, of which heat lysis was found to be efficient and convenient. The specificity for Vibrio vulnificus was tested using primers from vvhA and gyrB genes with 3 vibrio spp. and 7 other organisms which gave a specific 519 bp and 285 bp products for Vibrio vulnificus. The minimum detection level with pure Vibrio vulnificus culture was found to be 0.25 CFU/ml.
APW and Luria Bertani enrichment broths were evaluated for PCR compatibility, it revealed that APW broth was superior. Spiking studies were carried out by inoculating with pure culture, it revealed that 18h incubation was better than 12h.
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bacteria, biological phenomena, pcr, diseases, antibiotics, irrigation, sampling, genes, fruits, byproducts