Increased Sensitivity Loop-Amplification Using Activated Carbon and Bentonite for the Detection of Vibrio spp. in Fishery Products.

Abstract

The Vibrio genus has more than 100 species, of which 12 are considered pathogenic for humans; species most frequently isolated in food are V. cholerae, V. parahaemolyticus, V. vulnificus, and V. alginolyticus. The main problem of identifying them through conventional microbiological methods is the high costs and long-term diagnosis. Loop-mediated isothermal Amplification (LAMP) was used for the identification of V. cholerae, V. parahaemolyticus, V. vulnificus, and V. alginolyticus. Activated carbon with bentonite (ACB) was used to remove inhibitors, improving the sensitivity of the test; pH 6, 7, and 9 were tested, as well as different contact times between the sample and ACB. The LAMP sensitivity method was improved when activated carbon and bentonite were used to remove reaction inhibitors in samples of shrimp, fish, and oyster. LAMP sensitivity, negative predictive value, positive predictive value, and LAMP specificity were equal to or greater than 90% for all samples tested. The LAMP method could be an alternative to identify V. cholerae, V. parahaemolyticus, V. vulnificus, and V. alginolyticus in fishery products due to faster application, higher sensitivity, and lower costs than the standard method.