Abstract
Quantitative detection of V. vulnificus in shellfish via competitive PCR was herein developed. We designed a forward primer, a reverse primer and a hybrid forward primer based on the V. vulnificus specific hemolysin gene (vvh), which was used to amplify target DNA and an internal competitive PCR standard. The specificity of the primers for V. vulnificus was proven by gel electrophoresis. The detection sensitivity for V. vulnificus was 220 CFU per PCR reaction with cells from a pure culture and 270 CFU/g of tissue without enrichment. After 10 hours of enrichment at 37° C the minimum detection level was reduced to 7 CFU/g of tissue.
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