Detection of Salmonella species in foodstuffs.
Conventional methods to detect Salmonella spp. in foodstuffs may take up to 1 wk. Methods for pathogen detection are required. Real-time detection of Salmonella spp. will broaden our ability to screen large number of samples in a short time. This chapter describes a step-by-step procedure using an oligonucleotide probe that becomes fluorescent upon hybridization to the target DNA (Molecular Beacon; MB) in a real-time polymerase chain reaction (PCR) assay. The capability of the assay to detect Salmonella species from artificially inoculated fresh- and fresh-cut produce as well as ready-to-eat meats is demonstrated. The method uses internal positive and negative controls which enable researchers to detect false-negative PCR results. The procedure uses the buffered peptone water for the enrichment of Salmonella spp. and successfully detects the pathogen at low level of contamination (2-4 cells/25 g) in <24 h.
Bhagwat
Arvind A
AA
Produce Quality and Safety Laboratory, Henry A. Wallace Beltsville Agricultural Research Center, USDA, Beltsville, MD, USA.
Patel
Jitu
J
Chua
Trina
T
Chan
Audrey
A
Cruz
Saúl Ruiz
SR
Aguilar
Gustavo A González
GA
D015169:Colony Count, Microbial; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D008460:Meat / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012475:Salmonella / Q000302:isolation & purification*; D014675:Vegetables / Q000382:microbiology*