Use of quantitative real-time PCR for the detection of Salmonella spp. in fecal samples from horses at a veterinary teaching hospital

Nicola Pusterla, Barbara A Byrne, Emir Hodzic, Samantha Mapes, Spencer S. Jang, K G Magdesian

Research output: Contribution to journalArticle

29 Citations (Scopus)

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 24. h 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.

Original languageEnglish (US)
Pages (from-to)252-255
Number of pages4
JournalVeterinary Journal
Volume186
Issue number2
DOIs
StatePublished - Nov 2010

Fingerprint

Animal Hospitals
Feces
Teaching Hospitals
Salmonella
Horses
Real-Time Polymerase Chain Reaction
quantitative polymerase chain reaction
feces
horses
Selenious Acid
selenites
serotypes
sampling
assays
analytical specificity
microbial detection
cross infection
Bacterial Load
Salmonella Infections
organisms

Keywords

  • Feces
  • Horse
  • Microbiological culture
  • Quantitative real-time PCR
  • Salmonella spp.

ASJC Scopus subject areas

  • Animal Science and Zoology
  • veterinary(all)

Cite this

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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",
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 24. h 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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AU - Mapes, Samantha

AU - Jang, Spencer S.

AU - Magdesian, K G

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