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 language | English (US) |
|---|---|
| Pages (from-to) | 252-255 |
| Number of pages | 4 |
| Journal | Veterinary Journal |
| Volume | 186 |
| Issue number | 2 |
| DOIs | |
| State | Published - Nov 2010 |
Fingerprint
Keywords
- Feces
- Horse
- Microbiological culture
- Quantitative real-time PCR
- Salmonella spp.
ASJC Scopus subject areas
- Animal Science and Zoology
- veterinary(all)
Cite this
Use of quantitative real-time PCR for the detection of Salmonella spp. in fecal samples from horses at a veterinary teaching hospital. / Pusterla, Nicola; Byrne, Barbara A; Hodzic, Emir; Mapes, Samantha; Jang, Spencer S.; Magdesian, K G.
In: Veterinary Journal, Vol. 186, No. 2, 11.2010, p. 252-255.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Use of quantitative real-time PCR for the detection of Salmonella spp. in fecal samples from horses at a veterinary teaching hospital
AU - Pusterla, Nicola
AU - Byrne, Barbara A
AU - Hodzic, Emir
AU - Mapes, Samantha
AU - Jang, Spencer S.
AU - Magdesian, K G
PY - 2010/11
Y1 - 2010/11
N2 - 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.
AB - 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.
KW - Feces
KW - Horse
KW - Microbiological culture
KW - Quantitative real-time PCR
KW - Salmonella spp.
UR - http://www.scopus.com/inward/record.url?scp=78149396113&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=78149396113&partnerID=8YFLogxK
U2 - 10.1016/j.tvjl.2009.08.022
DO - 10.1016/j.tvjl.2009.08.022
M3 - Article
C2 - 19766027
AN - SCOPUS:78149396113
VL - 186
SP - 252
EP - 255
JO - Veterinary Journal
JF - Veterinary Journal
SN - 1090-0233
IS - 2
ER -