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  • Title: Application of gyrB targeted SYBR green based qPCR assay for the specific and rapid detection of Vibrio vulnificus in seafood.
    Author: D'Souza C, Kumar BK, Rai P, Deekshit VK, Karunasagar I.
    Journal: J Microbiol Methods; 2019 Nov; 166():105747. PubMed ID: 31639359.
    Abstract:
    A SYBR green based qPCR assay targeting a unique region of gyrB was developed for the detection of Vibrio vulnificus. The specificity of the assay was studied using V. vulnificus and other bacterial strains belonging to Vibrio and non-Vibrio species. The assay unambiguously distinguished V.vulnificus with a sensitivity of 101 CFU/mL in pure culture while 102CFU/g was detected in clam meat homogenate with an efficiency of ≥98%.The utility of the qPCR assay was validated with naturally incurred seafood samples, where 24 out of 59(40.67%) seafood samples tested positive for V. vulnificus after 6-8 h enrichment in APW-P broth. In contrast, conventional PCR could detect only 11 samples (18.64%). Our results showed that qPCR assay developed in this study could be used as a rapid method for screening seafood samples for the presence of V. vulnificus, as the assay can be completed within 9-12 h including the enrichment of seafood in APW-P broth. The gyrB targeted qPCR developed in this study can provide excellent results on the presence and load of V. vulnificus in naturally contaminated samples quickly and efficiently; thus it could find application as a routine test in the seafood industry for the analysis V. vulnificus.
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