title: Development of PCR assays for detection of Escherichia coli O157:H7 in meat products.
abstract: A multiplex polymerase chain reaction (PCR) procedure based on fliC(h7) and rfbE genes was developed for the detection of Escherichia coli O157:H7 in raw pork meat and ready-to-eat (RTE) meat products. Two different DNA extraction procedures were evaluated for application on meat products. MasterPure™ DNA Purification kit in combination with immunomagnetic separation was found to be the best method in a meat system. The optimized PCR included an enrichment step in brilliant green bile 2% broth at 37 °C. This method was applied to artificially inoculated meat and RTE meat products with different concentrations of E. coli O157:H7. The results indicate that the PCR assay developed could sensitively and specifically detect E. coli O157:H7 in raw pork meat and RTE meat products in approximately 10h, including a 6h enrichment step. Thus, this method could be proposed for screening E. coli O157:H7 in raw pork and RTE meat products.
authors: Food Hygiene and Safety, Faculty of Veterinary Science, University of Extremadura, Avda. de la Universidad s/n. 10003, Cáceres, Spain.
authors: Rubén
authors: R
authors: Gordillo
authors: Juan J
authors: JJ
authors: Córdoba
authors: María J
authors: MJ
authors: Andrade
authors: M Isabel
authors: MI
authors: Luque
authors: Mar
authors: M
authors: Rodríguez
mesh_terms: D000818:Animals; D004269:DNA, Bacterial / Q000302:isolation & purification; D019453:Escherichia coli O157 / Q000302:isolation & purification*; D005516:Food Microbiology / Q000379:methods*; D018189:Immunomagnetic Separation; D008461:Meat Products / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013552:Swine
chemical_list: D004269:DNA, Bacterial