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.
Abstract: A quantitative real-time polymerase chain reaction method (qPCR) was developed and tested for the detection of Taylorella equigenitalis. It was shown to have an analytical sensitivity of 5 colony-forming units (CFU) of T. equigenitalis when applied to the testing of culture swabs that mimicked field samples, and a high analytical specificity in not reacting to 8 other commensal bacterial species associated with horses. As designed, it could also differentiate specifically between T. equigenitalis and T. asinigenitalis. The qPCR was compared to standard culture in a study that included 45 swab samples from 6 horses (1 stallion, 5 mares) naturally infected with T. equigenitalis in Canada, 39 swab samples from 5 naturally infected stallions in Germany, and 311 swab samples from 87 culture negative horses in Canada. When the comparison was conducted on an individual sample swab basis, the qPCR had a statistical sensitivity and specificity of 100% and 96.4%, respectively, and 100% and 99.1% when the comparison was conducted on a sample set basis. A comparison was also made on 203 sample swabs from the 5 German stallions taken over a span of 4 to 9 mo following antibiotic treatment. The qPCR was found to be highly sensitive and at least as good as culture in detecting the presence of T. equigenitalis in post-treatment samples. The work demonstrates that the qPCR assay described here can potentially be used to detect the presence of T. equigenitalis directly from submitted sample swabs taken from infected horses and also for determining T. equigenitalis freedom following treatment. Une épreuve quantitative de réaction en chaîne par la polymérase en temps réel (qPCR) a été développée et testée pour la détection de Taylorella equigenitalis. L’épreuve a démontré une sensibilité analytique de 5 UFC de T. equigenitalis à partir de prélèvements de culture qui imitent les échantillons de terrain, et une spécificité analytique élevée puisqu’elle n’a pas réagi à 8 autres espèces bactériennes commensales associées aux chevaux. D’ailleurs, sa conception peut aussi faire la différenciation entre T. equigenitalis et T. asinigenitalis. La qPCR a été comparée à la méthode de culture classique de routine dans une étude qui a inclus 45 prélèvements provenant de 6 chevaux du Canada (1 étalon, 5 juments), 39 prélèvements de 5 étalons d’Allemagne, tous infectés naturellement avec T. equigenitalis, ainsi que 311 prélèvements provenant de 87 chevaux du Canada diagnostiqués négatifs par la méthode de détection par culture. Lorsque la comparaison a été faite pour les prélèvements individuels, la qPCR a démonstré une sensibilité et spécificité statistique de 100 % et 96,4 %, respectivement, et de 100 % et 99,1 %, pour la comparaison basée sur les séries de prélèvements. De plus, une comparaison a été faite à partir de 203 prélèvements de 5 étalons d’Allemagne ayant été pris dans un intervalle de 4 à 9 mois après un traitement aux antibiotiques. La qPCR s’est avérée hautement sensible et tout au moins aussi bonne que la méthode par culture pour la détection de T. equigenitalis dans les prélèvements post-traitement. Ce projet démontré que la qPCR, décrite ici, peut être utilisée pour la détection de la présence de T. equigenitalis directement des prélèvements de chevaux infectés ainsi que pour la confirmation de l’absence de T. equigenitalis après traitement.(Traduit par Ms. Émilie Falardeau).
Publication Date: 2015-07-02 PubMed ID: 26130847PubMed Central: PMC4445507
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- Journal Article
Summary
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This research compares the efficiency between qPCR and traditional culture methods in detecting Taylorella equigenitalis, a bacteria found in horses. It finds that qPCR is highly sensitive and, in many aspects, superior to the traditional culture method in detecting the presence of the bacteria in sample swabs from infected and treated horses.
Development and testing of qPCR
- The study first discussed the development of a quantitative real-time polymerase chain reaction method (qPCR) to detect Taylorella equigenitalis, a bacteria mainly found in horses.
- The qPCR technique displayed a high analytical sensitivity as it could detect as few as 5 colony-forming units of T. equigenitalis. This was determined using samples which closely represented field samples from horses.
- Furthermore, qPCR demonstrated a high level of specificity as it did not react to 8 other types of bacteria commonly found in horses, indicating its precise targeting capability.
- QPCR could also differentiate between T. equigenitalis and T. asinigenitalis, showing its ability to distinguish between similar species.
Comparison of qPCR and culture method
- The researchers then carried out a study to compare qPCR with the traditional culture method. This involved sampling swabs from 6 horses in Canada and 5 in Germany, all naturally infected with T. equigenitalis, as well as 311 swabs from 87 horses that tested negative for T. equigenitalis using the culture method.
- When each sample was examined individually, qPCR had a 100% sensitivity (ability to identify positive results) and 96.4% specificity (capacity to correctly identify negative results).
- When the analysis was carried out on a set of samples, the sensitivity and specificity of qPCR attained 100% and 99.1%, respectively.
qPCR’s efficiency in detecting post-treatment T. equigenitalis
- An additional comparison was made using 203 samples from the German horses over a span of 4 to 9 months following antibiotic treatment.
- In these post-treatment samples, qPCR was highly sensitive and at least as effective as the traditional culture in detecting the presence of T. equigenitalis.
- This suggests the potential utility of qPCR in confirming the elimination of T. equigenitalis after treatment.
Conclusion
- The research concluded that qPCR, as described in the paper, is effective in detecting the presence of T. equigenitalis and could potentially be used alongside or replace traditional culture methods.
- The method can be used directly from sample swabs taken from horses and is beneficial in confirming the absence of T. equigenitalis post-treatment.
Cite This Article
APA
Nadin-Davis S, Knowles MK, Burke T, Böse R, Devenish J.
(2015).
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, 79(3), 161-169.
Publication
Researcher Affiliations
- Animal Health Microbiology, CFIA/Ottawa Laboratory (Fallowfield), 3851 Fallowfield Road, P.O. Box 11300 Station H, Ottawa, Ontario (Nadin-Davis, Knowles, Burke, Devenish); Labor Dr. Böse GmbH, Carl-Zeiss-Str. 6, 31177 Harsum, Germany (Böse).
- Animal Health Microbiology, CFIA/Ottawa Laboratory (Fallowfield), 3851 Fallowfield Road, P.O. Box 11300 Station H, Ottawa, Ontario (Nadin-Davis, Knowles, Burke, Devenish); Labor Dr. Böse GmbH, Carl-Zeiss-Str. 6, 31177 Harsum, Germany (Böse).
- Animal Health Microbiology, CFIA/Ottawa Laboratory (Fallowfield), 3851 Fallowfield Road, P.O. Box 11300 Station H, Ottawa, Ontario (Nadin-Davis, Knowles, Burke, Devenish); Labor Dr. Böse GmbH, Carl-Zeiss-Str. 6, 31177 Harsum, Germany (Böse).
- Animal Health Microbiology, CFIA/Ottawa Laboratory (Fallowfield), 3851 Fallowfield Road, P.O. Box 11300 Station H, Ottawa, Ontario (Nadin-Davis, Knowles, Burke, Devenish); Labor Dr. Böse GmbH, Carl-Zeiss-Str. 6, 31177 Harsum, Germany (Böse).
- Animal Health Microbiology, CFIA/Ottawa Laboratory (Fallowfield), 3851 Fallowfield Road, P.O. Box 11300 Station H, Ottawa, Ontario (Nadin-Davis, Knowles, Burke, Devenish); Labor Dr. Böse GmbH, Carl-Zeiss-Str. 6, 31177 Harsum, Germany (Böse).
MeSH Terms
- Animals
- Bacteriological Techniques / veterinary
- Canada / epidemiology
- Female
- Germany / epidemiology
- Gram-Negative Bacterial Infections / diagnosis
- Gram-Negative Bacterial Infections / microbiology
- Gram-Negative Bacterial Infections / veterinary
- Horse Diseases / diagnosis
- Horse Diseases / microbiology
- Horses
- Male
- Sensitivity and Specificity
- Sexually Transmitted Diseases, Bacterial / diagnosis
- Sexually Transmitted Diseases, Bacterial / microbiology
- Sexually Transmitted Diseases, Bacterial / veterinary
- Taylorella equigenitalis / isolation & purification
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