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An investigation into the suitability of a commercial real-time PCR assay to screen for Taylorella equigenitalis in routine prebreeding equine genital swabs.
Abstract: Standard bacteriological methods for identifying Taylorella equigenitalis in cervical smears are time consuming. Therefore, a more rapid real-time PCR assay was evaluated for its suitability in screening swabs. Objective: To compare the results of a commercially available real-time PCR assay with routine microbiological culture for the identification of T. equigenitalis, the causative organism of contagious equine metritis, in equine genital swab samples, under 'field trial' conditions. Methods: Routine prebreeding genital swabs (n=2072) collected from Thoroughbred mares and stallions during 2009 were examined together with stored T. equigenitalis positive material. Swabs were cultured for T. equigenitalis using standard microbiological techniques. Bacterial lysates were isolated from the swabs and examined for the presence of a 16S DNA fragment of T. equigenitalis, using a commercial multiplex real-time PCR assay system. Results: There was complete concordance between positive and negative results obtained by the 2 methods. Real-time PCR also detected T. equigenitalis DNA from swabs that were negative using standard microbiological culture after 6 months' storage at +4 degrees C but from which T. equigenitalis had been isolated following collection. The sensitivities of real-time PCR and bacterial culture were both 10(-3) (equivalent to 3 colony-forming units). Conclusions: Routine bacterial culture of T. equigenitalis requires an incubation period of not less than 7 days before a conclusive negative result can be obtained, whereas bacterial extraction and real-time PCR assay can be completed in less than 6 h. The commercially-available PCR assay tested provided a rapid and reliable method for the identification of T. equigenitalis from equine genital swabs and could be usefully employed for the screening of mares and stallions for preseason Horserace Betting Levy Board (HBLB) Code of Practice and in other situations such as for bloodstock sales screening requirements, overcoming the current delays imposed by bacterial culture requirements. Its use could be quality assured by the existing HBLB biannual testing scheme for designated laboratories.
Publication Date: 2010-04-14 PubMed ID: 20383985DOI: 10.2746/042516409x474275Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article evaluates the effectiveness of a real-time PCR assay for rapidly identifying the bacterium Taylorella equigenitalis in equine genital swabs, in comparison with the time-consuming traditional microbiological culture methods. The researchers found the PCR assay to offer quicker and equally reliable results, suggesting its great potential for use in regular screenings before breeding and bloodstock sales.
Research Objective
- This research aimed to assess the viability of using a commercially available real-time polymerase chain reaction (PCR) assay as a fast and reliable approach for identifying T. equigenitalis in equine genital swab samples under practical conditions.
Methods
- The study used routine pre-breeding genital swabs gathered from thoroughbred horses in 2009, along with stored T. equigenitalis positive material.
- These swabs were tested for the presence of T. equigenitalis using both the traditional microbiological culture methods and the real-time PCR assay, which specifically looks for a fragment of the bacterium’s 16S DNA.
- Lastly, the researchers compared the results obtained from both methods.
Results
- The results showed perfect agreement between the PCR assay and the traditional culture method, with both having a sensitivity of 10(-3) or the equivalent of detecting 3 colony-forming units.
- Interestingly, the PCR test was able to identify T. equigenitalis DNA on swabs that were deemed negative by the culture method after six months of storage at relatively low temperatures.
Conclusion
- In conclusion, the researchers found the real-time PCR assay to be a highly efficient and reliable alternative to the time-intensive bacterial culture methods.
- This PCR-based method can be completed in under six hours and therefore, could drastically cut down the time needed for diagnosing whether horses carry the T. equigenitalis bacterium.
- The researchers suggest that this commercial PCR assay could be very useful in regular screenings for T. equigenitalis before breeding, sales of bloodstock, and conforming to the Horserace Betting Levy Board’s Code of Practice, all of which currently incur delays due to the lengthy nature of standard bacterial culture tests.
Cite This Article
APA
Ousey JC, Palmer L, Cash RS, Grimes KJ, Fletcher AP, Barrelet A, Foote AK, Manning FM, Ricketts SW.
(2010).
An investigation into the suitability of a commercial real-time PCR assay to screen for Taylorella equigenitalis in routine prebreeding equine genital swabs.
Equine Vet J, 41(9), 878-882.
https://doi.org/10.2746/042516409x474275 Publication
Researcher Affiliations
- Beaufort Cottage Laboratories, High Street, Newmarket, Suffolk CB8 8JS, UK.
MeSH Terms
- Animals
- Female
- Gram-Negative Bacterial Infections / microbiology
- Gram-Negative Bacterial Infections / prevention & control
- Gram-Negative Bacterial Infections / veterinary
- Horse Diseases / microbiology
- Horse Diseases / prevention & control
- Horses
- Male
- Polymerase Chain Reaction / methods
- Polymerase Chain Reaction / veterinary
- Sexually Transmitted Diseases, Bacterial / microbiology
- Sexually Transmitted Diseases, Bacterial / prevention & control
- Sexually Transmitted Diseases, Bacterial / veterinary
- Taylorella equigenitalis / isolation & purification
Citations
This article has been cited 3 times.- Wang Y, Li Z, Zuo Z, Gu X, Cai D, Hu J, Gu Y, Shen L, Gou L, Zhang K, Ma X. Establishment and application of multiplex PCR method for detection of Trichophyton verrucosum, Microsporum canis, and Trichophyton mentagrophytes from cattle. Front Vet Sci 2025;12:1546586.
- Mawhinney I, Bollard A. Enhanced detection of Taylorella equigenitalis by qPCR using 'Dry' swabs. J Equine Sci 2023 Mar;34(1):7-12.
- Nadin-Davis S, Knowles MK, Burke T, Böse R, Devenish J. Comparison of culture versus quantitative real-time polymerase chain reaction for the detection of Taylorella equigenitalis in field samples from naturally infected horses in Canada and Germany. Can J Vet Res 2015 Jul;79(3):161-9.
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