Comparison of polymerase chain reaction and microbiological culture for detection of salmonellae in equine feces and environmental samples.
Abstract: To compare the sensitivity of polymerase chain reaction (PCR) with microbiological culture for detecting salmonellae in equine fecal samples and equine environmental swab specimens. Methods: Samples and specimens were tested by PCR and microbiological culture. Methods: A fecal sample from each of 152 horses admitted consecutively to the clinic for evaluation by the outpatient service, 282 fecal samples from 110 hospitalized horses that had been submitted to the clinical microbiology laboratory, and 313 environmental swab specimens were examined. Methods: Each sample and specimen in the study was tested, using PCR and microbiological culture. Results of PCR and culture were compared. Results: Significantly (P < 0.001) more fecal samples were positive by PCR than by microbiological culture. 26 of 152 (17.1%) fecal samples collected from horses admitted by the outpatient service were positive by PCR and none was positive by culture. 71 of 110 hospitalized horses were identified as positive by PCR, compared with 11 horses identified as positive by culture. All culture-positive horses were positive by PCR. Of the 11 culture-positive horses, 10 (90.9%) were identified as PCR positive after testing of the first sample submitted, compared with 7 (63.6%) by culture. All PCR-positive horses were detected after a total of 3 samples/horse were submitted, whereas as many as 5 samples/horse was required to identify all culture-positive horses. 8 of 313 environmental specimens were positive by PCR, and none was positive by culture. Conclusions: The PCR method reported here was more sensitive, more rapid, and required submission of fewer samples or specimens than did microbiological culture for detecting salmonellae.
Publication Date: 1996-06-01 PubMed ID: 8725799
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research investigates the efficiency of the polymerase chain reaction (PCR) method versus traditional microbiological culture in detecting the presence of salmonellae in horse feces and environmental samples. The study indicates PCR is more reliable, faster, and requires fewer samples compared to microbiological culture.
Methods Used
- The research included samples from different sources: 152 fecal samples from horses admitted for evaluation, 282 samples from 110 hospitalized horses, and 313 environmental swabs.
- Both PCR and microbiological culture testing were conducted on each sample to compare their efficiency in the detection of salmonellae.
Research Findings
- The research findings indicated that the PCR method was significantly more successful in detecting salmonellae in fecal samples than the standard microbiological culture.
- PCR detected salmonellae in 17.1% of samples from outpatient service horses, while the culture method detected none.
- Of the 110 hospitalized horses, PCR identified 71 positive cases while the culture method recognized only 11.
- All samples that tested positive by culture also tested positive by PCR, suggesting that PCR is equally, if not more, reliable.
Efficiency Comparison
- PCR identified 90.9% of positive cases in the first sample test, while the culture method detected only 63.6%.
- All PCR-positive cases were identified within the first three sample tests, while the culture method required up to five sample tests to identify all positive cases.
- PCR detected salmonellae in eight out of 313 environmental specimens, while the culture method detected none.
Conclusions
- The research results point out that PCR is not only more sensitive in detecting salmonellae but also faster and requires fewer samples compared to microbiological culture.
- This suggests that the application of PCR could help in the effective and swift detection of salmonellae, thereby contributing to faster and more effective treatments.
Cite This Article
APA
Cohen ND, Martin LJ, Simpson RB, Wallis DE, Neibergs HL.
(1996).
Comparison of polymerase chain reaction and microbiological culture for detection of salmonellae in equine feces and environmental samples.
Am J Vet Res, 57(6), 780-786.
Publication
Researcher Affiliations
- Department of Large Animal Medicine and Surgery, College of Veterinary Medicine, Texas A&M University, College Station 77843, USA.
MeSH Terms
- Animals
- Base Sequence
- DNA Primers / chemistry
- DNA, Bacterial / analysis
- DNA, Bacterial / chemistry
- DNA, Bacterial / genetics
- Environmental Microbiology
- Feces / microbiology
- Female
- Horse Diseases / diagnosis
- Horse Diseases / microbiology
- Horses
- Male
- Microbiological Techniques / standards
- Microbiological Techniques / veterinary
- Molecular Sequence Data
- Polymerase Chain Reaction / standards
- Polymerase Chain Reaction / veterinary
- Salmonella / genetics
- Salmonella / isolation & purification
- Salmonella Infections, Animal / diagnosis
- Salmonella Infections, Animal / microbiology
- Sensitivity and Specificity
- Time Factors
Citations
This article has been cited 15 times.- Amory H, Cesarini C, De Maré L, Loublier C, Moula N, Detilleux J, Saulmont M, Garigliany MM, Lecoq L. Relationship between the Cycle Threshold Value (Ct) of a Salmonella spp. qPCR Performed on Feces and Clinical Signs and Outcome in Horses. Microorganisms 2023 Jul 30;11(8).
- Ownagh A, Etemadi N, Khademi P, Tajik H. Identification of Salmonella carriers by amplification of FimA, Stn and InvA genes and bacterial culture methods in fecal samples of buffalo. Vet Res Forum 2023;14(1):21-28.
- Uzal FA, Arroyo LG, Navarro MA, Gomez DE, Asín J, Henderson E. Bacterial and viral enterocolitis in horses: a review. J Vet Diagn Invest 2022 May;34(3):354-375.
- Knox A, Beddoe T. Isothermal Nucleic Acid Amplification Technologies for the Detection of Equine Viral Pathogens. Animals (Basel) 2021 Jul 20;11(7).
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- Shaw SD, Stämpfli H. Diagnosis and Treatment of Undifferentiated and Infectious Acute Diarrhea in the Adult Horse. Vet Clin North Am Equine Pract 2018 Apr;34(1):39-53.
- Goyal SM, Anantharaman S, Ramakrishnan MA, Sajja S, Kim SW, Stanley NJ, Farnsworth JE, Kuehn TH, Raynor PC. Detection of viruses in used ventilation filters from two large public buildings. Am J Infect Control 2011 Sep;39(7):e30-8.
- García R, Baelum J, Fredslund L, Santorum P, Jacobsen CS. Influence of temperature and predation on survival of Salmonella enterica serovar Typhimurium and expression of invA in soil and manure-amended soil. Appl Environ Microbiol 2010 Aug;76(15):5025-31.
- Alexander KA, Warnick LD, Wiedmann M. Antimicrobial resistant Salmonella in dairy cattle in the United States. Vet Res Commun 2009 Mar;33(3):191-209.
- Sibley J, Yue B, Huang F, Harding J, Kingdon J, Chirino-Trejo M, Appleyard GD. Comparison of bacterial enriched-broth culture, enzyme linked immunosorbent assay, and broth culture-polymerase chain reaction techniques for identifying asymptomatic infections with Salmonella in swine. Can J Vet Res 2003 Jul;67(3):219-24.
- Gentry-Weeks C, Hutcheson HJ, Kim LM, Bolte D, Traub-Dargatz J, Morley P, Powers B, Jessen M. Identification of two phylogenetically related organisms from feces by PCR for detection of Salmonella spp. J Clin Microbiol 2002 Apr;40(4):1487-92.
- Feder I, Nietfeld JC, Galland J, Yeary T, Sargeant JM, Oberst R, Tamplin ML, Luchansky JB. Comparison of cultivation and PCR-hybridization for detection of Salmonella in porcine fecal and water samples. J Clin Microbiol 2001 Jul;39(7):2477-84.
- Carli KT, Unal CB, Caner V, Eyigor A. Detection of salmonellae in chicken feces by a combination of tetrathionate broth enrichment, capillary PCR, and capillary gel electrophoresis. J Clin Microbiol 2001 May;39(5):1871-6.
- Navarre CB, Belknap EB, Rowe SE. Differentiation of gastrointestinal diseases of calves. Vet Clin North Am Food Anim Pract 2000 Mar;16(1):37-57.
- Ravary B, Fecteau G, Higgins R, Paré J, Lavoie JP. [Prevalence of infections caused by Salmonella spp. in cattle and horses at the Veterinary Teaching Hospital of the Faculty of Veterinary Medicine of the University of Montreal]. Can Vet J 1998 Sep;39(9):566-72.
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