Evaluation of a newly developed real-time PCR for the detection of Taylorella equigenitalis and discrimination from T. asinigenitalis.
A 'culture-LightCycler PCR' assay has been developed for the detection of Taylorella equigenitalis, the causative agent of contagious equine metritis (CEM) in horses. The primers and hybridisation probes were derived from the 16S rDNA sequence. Their specificity was determined in two closely related organisms and six commensal bacteria of the genital tract. The assay was specific for T. equigenitalis and discriminates T. asinigenitalis isolates. It also avoids misidentifications of morphologically and phenotypically similar organisms. The sensitivity was evaluated in comparison to a standard bacteriological culture method. It detected T. equigenitalis in 10 of 52 samples that had not been identified bacteriologically. The results indicated that the assay had a greater sensitivity. This is the first real-time PCR for the detection of T. equigenitalis and avoids PCR carry-over contamination. The 'culture-LightCycler PCR' assay is specific, sensitive and reproducible, and can be used effectively for the detection of T. equigenitalis infections.
Premanandh
J
J
Central Veterinary Research Laboratory, P.O. Box 597, Dubai, United Arab Emirates.
George
L V
LV
Wernery
U
U
Sasse
J
J
D000818:Animals; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004716:Endometritis / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D005260:Female; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D006734:Horse Diseases / Q000382:microbiology*; D006736:Horses; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020625:Taylorella equigenitalis / Q000235:genetics* / Q000302:isolation & purification
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S