title: Rapid detection of Salmonella from hydrodynamic pressure-treated poultry using molecular beacon real-time PCR.
abstract: A real-time polymerase chain reaction (PCR) assay was evaluated to detect Salmonella in hydrodynamic pressure (HDP)-treated chicken using molecular beacon probes available as a commercial kit (iQ-Check, Bio-Rad Laboratories). The sensitivity and accuracy of the assay were compared with the conventional USDA microbiological procedure using artificially contaminated minced chicken. Chicken fillets were irradiated at 10 kGy to completely destroy any naturally occurring Salmonella. These fillets were minced and inoculated with as low as 2+/-1 cfu of S. typhimurium per 25 g chicken. The minced chicken samples were vacuum packed in multi-layer barrier bags, heat shrunk, and treated with HDP. Results showed that all inoculated samples (n=36) were detected by the PCR assay and conventional USDA procedure. Similarly, all uninoculated controls (n=11) were negative by both PCR assay and USDA procedure. As few as 2+/-1 cfu could be detected from 25 g HDP-treated chicken following 16-18 h enrichment in buffered peptone water. Real-time PCR proved to be an effective method for Salmonella detection in HDP-treated chicken with high sensitivity and more importantly, a rapid and high-throughput detection in 18 h, compared to 3-8 days for the conventional microbiological methods. HDP treatment, which has been reported to reduce spoilage bacteria in various meats, was unable to kill pathogenic Salmonella in minced chicken.
authors.affiliation: Food Technology and Safety Laboratory, Agricultural Research Service, USDA, Bldg. 201, 10300 Baltimore Avenue, Beltsville, MD 20705-2350, USA. jpatel@anri.barc.usda.gov
authors.forename: J R
authors.initials: JR
authors.lastname: Patel
authors.forename: A A
authors.initials: AA
authors.lastname: Bhagwat
authors.forename: G C
authors.initials: GC
authors.lastname: Sanglay
authors.forename: M B
authors.initials: MB
authors.lastname: Solomon
mesh_terms: D000818:Animals; D002645:Chickens; D015169:Colony Count, Microbial; D005506:Food Contamination / Q000032:analysis*; D005514:Food Irradiation; D005516:Food Microbiology; D018857:Food Packaging / Q000379:methods; D006874:Hydrostatic Pressure; D016133:Polymerase Chain Reaction / Q000379:methods*; D011202:Poultry Products / Q000382:microbiology*; D015203:Reproducibility of Results; D012475:Salmonella / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013997:Time Factors