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Validation of an Easy Handling Sample Preparation and Triplex Real Time PCR for Rapid Detection of T. equigenitalis and Other Organisms Associated with Endometritis in Mares

Validation of an Easy Handling Sample Preparation and Triplex Real Time PCR for Rapid Detection of T. equigenitalis and Other Organisms Associated with Endometritis in Mares

Authors :
Albertine Léon
Yann Versmisse
Léa Despois
Patrice Gracieux
Béatrice Blanchard
Sophie Castagnet
LABÉO, Pôle d’analyses et de recherche de Normandie (LABÉO)
Unité de Recherche Risques Microbiens (U2RM)
Université de Caen Normandie (UNICAEN)
Normandie Université (NU)-Normandie Université (NU)
Adiagene
Source :
Journal of Equine Veterinary Science, Journal of Equine Veterinary Science, WB Saunders, 2020, 94, pp.103241. ⟨10.1016/j.jevs.2020.103241⟩
Publication Year :
2020
Publisher :
Elsevier BV, 2020.

Abstract

International audience; Isolation and identification of Taylorella equigenitalis, the causative agent of contagious equine metritis, by bacteriology is laborious and does not permit differentiation from the other member of the genus, Taylorella asinigenitalis. Moreover, other organisms such as Klebsiella pneumoniae and Pseudomonas aeruginosa can also cause endometritis in mares and warrant diagnostic detection. Our objectives were to develop a rapid preparation method for field swab samples and to validate this protocol using new multiplex real-time polymerase chain reaction (rtPCR) detection tools for identification of these four pathogens. The complete analytical process from sample preparation to PCR analysis was then evaluated against bacteriology, the World Organisation for Health's (OIE) gold standard method for T. equigenitalis and commonly used for the other three pathogens. The diagnostic sensitivity and specificity of this method, which used direct lysis and a multiplex rtPCR, were 100% and >92%, respectively. This study provided a simple-to-use method for prebreeding screening of mares and stallions.

Details

ISSN :
07370806
Volume :
94
Database :
OpenAIRE
Journal :
Journal of Equine Veterinary Science
Accession number :
edsair.doi.dedup.....9387a32544e272f38597a38f1e7ad26c
Full Text :
https://doi.org/10.1016/j.jevs.2020.103241