Rapid PCR detection of Salmonella in horse faecal samples.
Abstract: A rapid polymerase chain reaction (PCR) assay was developed for detecting Salmonella in faeces of horses and assessed on samples from horses admitted to a veterinary hospital. Direct detection was achieved by amplification of part of ompC after extraction of DNA from faeces using a spin column method to reduce the amount of inhibitory substances in samples. An internal positive control was included to detect false negative results. While the sensitivity of the PCR assay was less than culture when assessed on faeces inoculated with Salmonella, its sensitivity on faecal samples obtained from horses was much greater than culture. Salmonella DNA was detected in 40% of faecal samples using the PCR assay while Salmonella were cultured from only 2% of the samples. The PCR assay has potential for use in either routine diagnosis or for detection of the carrier status in animals.
Publication Date: 2001-03-07 PubMed ID: 11230929DOI: 10.1016/s0378-1135(00)00340-0Google Scholar: Lookup
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Summary
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The research article focuses on the development and application of a polymerase chain reaction (PCR) assay for detecting Salmonella in horse feces, and its comparison with traditional culture methods.
PCR Assay Development
- The researchers developed a rapid Polymerase Chain Reaction (PCR) assay to detect Salmonella in the feces of horses. The PCR amplifies a part of the ompC gene which is specific to Salmonella.
- The DNA for this process was extracted from the feces using a method called ‘spin column’, which significantly reduces the amount of inhibitory substances usually present in fecal samples.
- An internal positive control was included in the PCR assay to ensure accuracy and allow the detection of any ‘false negatives’.
Comparative Analysis Between PCR Assay and Culture Method
- The research then compares the sensitivity (ability to accurately identify positive samples) of the PCR assay with the traditional culture method.
- For feces samples that were artificially inoculated with Salmonella, the sensitivity of the culture method was higher compared to the PCR assay.
- However, in the case of naturally obtained fecal samples from horses, the PCR assay proved to have a significantly higher sensitivity compared to the culture method.
- Only 2% of these natural samples tested positive for Salmonella using the culture method, whereas 40% were found to be positive when tested with the PCR assay.
Implications of The Study
- The findings of this study suggest that PCR assays could be a more effective tool for detecting Salmonella in fecal samples compared to traditional culture methods.
- Despite the lower sensitivity in artificially inoculated samples, the PCR’s superior performance in natural samples demonstrates its potential for use in routine diagnosis or detection of carrier status in animals.
Cite This Article
APA
Amavisit P, Browning GF, Lightfoot D, Church S, Anderson GA, Whithear KG, Markham PF.
(2001).
Rapid PCR detection of Salmonella in horse faecal samples.
Vet Microbiol, 79(1), 63-74.
https://doi.org/10.1016/s0378-1135(00)00340-0 Publication
Researcher Affiliations
- Faculty of Veterinary Science, The University of Melbourne, Vic. 3010, Parkville, Australia.
MeSH Terms
- Animals
- Colony Count, Microbial / veterinary
- Feces / microbiology
- Horses / microbiology
- Polymerase Chain Reaction / methods
- Polymerase Chain Reaction / veterinary
- Porins / genetics
- Salmonella / isolation & purification
- Sensitivity and Specificity
Citations
This article has been cited 6 times.- Martínez-Pérez P, Hyndman TH, Fleming PA. Salmonella in Free-Ranging Quokkas (Setonix brachyurus) from Rottnest Island and the Mainland of Western Australia.. Animals (Basel) 2020 Mar 31;10(4).
- Milton AAP, Agarwal RK, Priya GB, Athira CK, Saminathan M, Reddy A, Aravind M, Kumar A. Occurrence, antimicrobial susceptibility patterns and genotypic relatedness of Salmonella spp. isolates from captive wildlife, their caretakers, feed and water in India.. Epidemiol Infect 2018 Sep;146(12):1543-1549.
- Behera P, Kutty VH, Kumar A, Sharma B. Changing the Codon Usage of hfq Gene has Profound Effect on Phenotype and Pathogenicity of Salmonella Typhimurium.. Curr Microbiol 2016 Mar;72(3):288-96.
- Hayashi M, Natori T, Kubota-Hayashi S, Miyata M, Ohkusu K, Kawamoto K, Kurazono H, Makino S, Ezaki T. A new protocol to detect multiple foodborne pathogens with PCR dipstick DNA chromatography after a six-hour enrichment culture in a broad-range food pathogen enrichment broth.. Biomed Res Int 2013;2013:295050.
- Slovis NM, Elam J, Estrada M, Leutenegger CM. Infectious agents associated with diarrhoea in neonatal foals in central Kentucky: a comprehensive molecular study.. Equine Vet J 2014 May;46(3):311-6.
- Wolffs PF, Glencross K, Thibaudeau R, Griffiths MW. Direct quantitation and detection of salmonellae in biological samples without enrichment, using two-step filtration and real-time PCR.. Appl Environ Microbiol 2006 Jun;72(6):3896-900.
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