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

Hemant Naikare; Daniela Bruno; Debabrata Mahapatra; Alesia Reinisch; Russell Raleigh; Robert Sprowls

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

Hemant Naikare, Daniela Bruno, Debabrata Mahapatra, Alesia Reinisch, Russell Raleigh, Robert Sprowls

Research output: Contribution to journal › Article › peer-review

16 Scopus citations

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.

Original language	English (US)
Pages (from-to)	32-42
Number of pages	11
Journal	Veterinary Sciences
Volume	2
Issue number	1
DOIs	https://doi.org/10.3390/vetsci2010032
State	Published - Mar 1 2015
Externally published	Yes

Bibliographical note

Publisher Copyright:
© 2015 by the authors.

Keywords

Cattle
Mycoplasma bovis
Taqman real-time PCR
uvrC gene

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10.3390/vetsci2010032

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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. / Naikare, Hemant; Bruno, Daniela; Mahapatra, Debabrata et al.
In: Veterinary Sciences, Vol. 2, No. 1, 01.03.2015, p. 32-42.

Research output: Contribution to journal › Article › peer-review

Naikare, H, Bruno, D, Mahapatra, D, Reinisch, A, Raleigh, R & Sprowls, R 2015, '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', Veterinary Sciences, vol. 2, no. 1, pp. 32-42. https://doi.org/10.3390/vetsci2010032

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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.",

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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

Abstract

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Keywords

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