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Detection and differentiation of bovine herpesvirus 1 and 5 using a multiplex real-time polymerase chain reaction.

Diallo, I.S. and Corney, B.G. and Rodwell, B.J. (2011) Detection and differentiation of bovine herpesvirus 1 and 5 using a multiplex real-time polymerase chain reaction. Journal of Virological Methods, 175 (1). pp. 46-52.

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Article Link(s): http://dx.doi.org/10.1016/j.jviromet.2011.04.013

Publisher URL: http://www.elsevier.com

Abstract

A multiplex real-time PCR was developed for the detection and differentiation of two closely related bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5). The multiplex real-time PCR combines a duplex real-time PCR that targets the DNA polymerase gene of BoHV-1 and BoHV-5 and a real-time PCR targeting mitochondrial DNA, as a house-keeping gene, described previously by Cawthraw et al. (2009). The assay correctly identified 22 BoHV-1 and six BoHV-5 isolates from the Biosecurity Sciences Laboratory virus collection. BoHV-1 and BoHV-5 were also correctly identified when incorporated in spiked semen and brain tissue samples. The detection limits of the duplex assay were 10 copies of BoHV-1 and 45 copies of BoHV-5. The multiplex real-time PCR had reaction efficiencies of 1.04 for BoHV-1 and 1.08 for BoHV-5. Standard curves relating Ct value to template copy number had correlation coefficients of 0.989 for BoHV-1 and 0.978 for BoHV-5. The assay specificity was demonstrated by testing bacterial and viral DNA from pathogens commonly isolated from bovine respiratory and reproductive tracts. The validated multiplex real-time PCR was used to detect and differentiate BoHV-1 and BoHV-5 in bovine clinical samples with known histories.

Item Type:Article
Business groups:Biosecurity Queensland
Additional Information:Crown Copyright © 2011
Keywords:Accuracy; assays; brain; diagnosis; genes; livestock; mitochondrial DNA; real time PCR; semen; viral diseases; multiplex real-time PCR; Bovine herpesvirus 1; Bovine herpesvirus 5; DNA viruses.
Subjects:Science > Biology > Genetics
Veterinary medicine > Diseases of special classes of animals
Deposited On:24 Nov 2011 06:23
Last Modified:05 Dec 2011 02:53

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