title: lolB gene, a valid alternative for qPCR detection of Vibrio cholerae in food and environmental samples.
abstract: 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.
authors.affiliation: ANFACO-CECOPESCA, Food Safety Division, Campus Univ. 16, 36310 Vigo PO, Spain; Microbiology and Toxins Area, ANFACO-CECOPESCA, Campus Univ. 16, 36310 Vigo PO, Spain.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Garrido-Maestu
authors.affiliation: ANFACO-CECOPESCA, Food Safety Division, Campus Univ. 16, 36310 Vigo PO, Spain.
authors.forename: María-José
authors.initials: MJ
authors.lastname: Chapela
authors.affiliation: ANFACO-CECOPESCA, Food Safety Division, Campus Univ. 16, 36310 Vigo PO, Spain.
authors.forename: Juan M
authors.initials: JM
authors.lastname: Vieites
authors.affiliation: ANFACO-CECOPESCA, Food Safety Division, Campus Univ. 16, 36310 Vigo PO, Spain. Electronic address: agcabado@anfaco.es.
authors.forename: Ana G
authors.initials: AG
authors.lastname: Cabado
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D015373:Bacterial Typing Techniques / Q000379:methods*; D004783:Environmental Microbiology; D005516:Food Microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D001426:Bacterial Proteins
keywords: Multiplex qPCR; Validation; Vibrio cholerae; ctxA; lolB; ompW