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Eight-hour PCR-based procedure for the detection of Salmonella in raw oysters
M Dolores Vázquez-Novelle1, Antonio J Pazos, Marcelina Abad
1Laboratorio de Biología Molecular y del Desarrollo, Departamento de Bioquímica y Biología Molecular, Instituto de Acuicultura, Universidad de Santiago de Compostela, 15782-Santiago de Compostela, Spain.
Abstract:
The aim of this study was to evaluate the ability of a nested PCR system to detect Salmonella senftenberg in raw oysters. The specific primers of the PCR were derived from the invA gene sequence, essential for Salmonella invasiveness into epithelial cells. First, for the extraction of DNA, four methods (guanidine isothiocyanate, E.Z.N.A. Mollusc Kit, Chelex-100, and lysis with detergents) were compared. A nested PCR method combined with 3.5 h pre-enrichment in buffered peptone water (BPW) and DNA extraction by the resin Chelex-100 is proposed for the detection of S. senftenberg in oyster samples. The detection limit of the method is less than 0.1 CFU/ml (<1 CFU/g of oyster). This procedure is shown to be an excellent tool for the sensitive detection of S. senftenberg from naturally contaminated oysters, with results being obtained within 8 h.
