Development and Evaluation of a Novel Taqman Real-Time PCR Assay for Rapid Detection of : Comparison of Assay Performance with a Conventional PCR Assay and Another Taqman Real-Time PCR Assay.

The objective of this study was to develop and validate a Taqman real-time PCR assay for the detection of (). Unique primers targeting the highly conserved house-keeping gene () were designed and the probe sequence was derived from a previously published microarray study. There was 100% agreement in the outcome between our assay and the other two published assays for detection. The analytical limit of detection of our assay is 83 copies of the gene. This assay was validated on a total of 214 bovine clinical specimens that were submitted to the Texas A&M Veterinary Medical Diagnostic Laboratory (TVMDL), Texas, USA. The specificity of the assay was assessed to be 100% since no cross-reactivity occurred with 22 other bacterial and other species. We conclude that the gene serves as a good and reliable diagnostic marker for the accurate and rapid detection of from a wider variety of specimen matrices.