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  • Title: Use of quantitative real-time PCR for the detection of Salmonella spp. in fecal samples from horses at a veterinary teaching hospital.
    Author: Pusterla N, Byrne BA, Hodzic E, Mapes S, Jang SS, Magdesian KG.
    Journal: Vet J; 2010 Nov; 186(2):252-5. PubMed ID: 19766027.
    Abstract:
    A quantitative real-time (RT)-PCR assay was developed to detect Salmonella spp. in the feces of 911 equine species admitted to a veterinary hospital. Fresh feces and feces enriched for 24h in selenite broth were assessed by conventional culture and by RT-PCR targeting the Salmonella invA gene. The detection limit for the RT-PCR assay was 3 and 10 organisms, respectively, when spiked samples were purified from selenite broth and feces. The analytical specificity was 100% based on the detection of a panel of 40 salmonella serotypes from five serogroups and the lack of cross-reactivity with non-related micro-organisms. Although Salmonella spp. were not cultured from fresh feces, the organism was cultured from 6/911 (0.6%) of broth-enriched samples. The bacterial load in enriched samples varied from 3 to 861,037 salmonella invA gene copies/μL DNA. The RT-PCR assay had an overall relative accuracy of 98%, a relative sensitivity of 100% and a relative specificity of 98%, when compared to conventional culture. The judicious use of such a RT-PCR method has the potential to reduce the risk of nosocomial infections such as salmonellosis through the provision of highly accurate and rapid pathogen detection.
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