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Vet. Sci. 2015, 2(1), 32-42; doi:10.3390/vetsci2010032

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

Texas A&M Veterinary Medical Diagnostic Laboratory, 6610 Amarillo Blvd West, Amarillo, TX 79106, USA
Some of the data included in this manuscript was presented at the 55th American Association of Veterinary Laboratory Diagnosticians (AAVLD) Annual Meeting, Greensboro, NC, USA, 18–24 October 2012.
These authors contributed equally to this work.
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Author to whom correspondence should be addressed.
Academic Editors: Duncan C. Ferguson and Margarethe Hoenig
Received: 27 December 2014 / Revised: 14 February 2015 / Accepted: 6 March 2015 / Published: 16 March 2015
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Abstract

The objective of this study was to develop and validate a Taqman real-time PCR assay for the detection of Mycoplasma bovis (M. bovis). Unique primers targeting the highly conserved house-keeping gene (uvrC) 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 M. bovis detection. The analytical limit of detection of our assay is 83 copies of the uvrC 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 Mycoplasma species. We conclude that the uvrC gene serves as a good and reliable diagnostic marker for the accurate and rapid detection of M. bovis from a wider variety of specimen matrices. View Full-Text
Keywords: Mycoplasma bovis; cattle; Taqman real-time PCR; uvrC gene Mycoplasma bovis; cattle; Taqman real-time PCR; uvrC gene
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Naikare, H.; Bruno, D.; Mahapatra, D.; Reinisch, A.; Raleigh, R.; Sprowls, R. Development and Evaluation of a Novel Taqman Real-Time PCR Assay for Rapid Detection of Mycoplasma bovis: Comparison of Assay Performance with a Conventional PCR Assay and Another Taqman Real-Time PCR Assay. Vet. Sci. 2015, 2, 32-42.

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