Journal
FOOD MICROBIOLOGY
Volume 46, Issue -, Pages 535-540Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.fm.2014.09.012
Keywords
Vibrio cholerae; Multiplex qPCR; Validation; lolB; ompW; ctxA
Funding
- Secretary General for the Sea of the Spanish Ministry of Agricultural, Land and Marine Resources (MARM) [ARM/1193/2009]
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In recent years a new genetic target for Vibrio cholerae detection has been reported, but its application was limited to clinical samples. This target, lolB, has never been applied to either food or environmental samples. In the present study the development, as well as the evaluation and pre-validation, of a real-time PCR method based on lolB, is described. The method included a newly designed hydrolysis probe to enhance its specificity. After comparison against other molecular and traditional methods, similar results were obtained regarding relative sensitivity, relative specificity, relative accuracy, positive and negative predictive values and index kappa of concordance (all higher than 91%), as well as a very low limit of detection (2 cfu/25 g). Additionally, after the analysis of more than 160 different food and environmental samples, its applicability in the food industry was completely demonstrated. (C) 2014 Elsevier Ltd. All rights reserved.
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