Two‐stage Nested PCR Effectiveness for Direct Detection of <i>Vibrio vulnificus</i> in Natural Samples

Abstract

ABSTRCT The nested primers designed to amplify a 222‐base pair portion of the hemolysin gene, vvhA , were specific for all V. vulnificus strains tested. The nested PCR amplification, coupled with direct extraction of template DNA, revealed improved sensitivity sufficient for detection of 1 to 10 CFU V. vulnificus in 1 mL of seafood homogenates, and eliminated the need for enrichment culturing. Thereby, the nested PCR method achieved a broader applicability, making it effective for extensive use in identification of the pathogen in natural samples such as raw seafoods, seawater and sediments.