Development of a real time PCR for the detection of Taylorella equigenitalis directly from genital swabs and discrimination from Taylorella asinigenitalis

Contagious equine metritis in Portugal: A retrospective report of the first outbreak in the country and recent contagious equine metritis test results

“…Three different "chocolate" blood agar media were used, one with no inhibitors (inhibitors may prevent the isolation of some strains of T. equigenitalis), one containing streptomycin sulphate (200 µg/ml) to allow for the differentiation of Streptomycin-sensitive or resistant biotypes, and the third medium according to , a medium with inhibitors, containing 1 µg/ml trimethoprim, 5µg/ml clindamycin and 5 µg/ml amphotericin B, which allows the growth of both streptomycin resistant and sensitive biotypes and is particularly useful to suppress growth of commensal bacteria and inhibit fungal growth (OIE, 2012 Plates were incubated at 37° C in a 5% (v/v) CO2 incubator and an incubation period of 10 days with no growth of suspect colonies was allowed before considering specimens negative for T. equigenitalis. The first T. equigenitalis strain isolated in Portugal (strain 18456) (Rocha, 2014) was sent for confirmation to the OIE reference laboratory for CEM (VLA, Bury St. Edmunds, UK), which was done both by observation of the phenotypic characteristics and by real-time PCR (Wakeley et al, 2006). All other 6 isolates obtained during the outbreak investigation, from the first 2 positive mares (M1 and M2) and the next 3 positive mares and 1 stallion (M3, M4, M5 and St2), besides the phenotypic and monoclonal antibodies identification as T. equigenitallis, were further tested at the INIAV according to the method described by Anzai et al (1999), a single step PCR test in a 2% agarose gel, using the primer set P1-N2, which amplifies a 445-bp DNA fragment of T. equigenitallis, allowing for its specific detection (Fig.…”

Infectious Diseases in Breeding Stallions

“…10 Additionally, a PCR method has recently been validated to test directly from swabs. 11 Control is predominantly through testing of imported horses to prevent spread of disease. Even with comprehensive standardized testing protocols, potential for transmission has been demonstrated to occur.…”

Detection of Taylorella equigenitalis and Taylorella asinigenitalis in horses in Croatia as a result of small scale survey

“…however, due to their similar growth requirements, morphology and biochemical characteristics, it is important to differentiate T. asinigenitalis from T. equigenitalis. This can be achieved by using molecular methods, with species specific primers, preferably qPCR, which also increases test sensitivity and significantly shortens turnaround time (WakeLeY et al, 2006;MaY et al, 2016).…”