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Sensitive and specific detection of proviral bovine leukemia virus by 5' Taq nuclease PCR using a 3' minor groove binder fluorogenic probe

Lew, A.E. and Bock, R.E. and Molloy, J.B. and Minchin, C.M. and Robinson, S.J. and Steer, P. (2004) Sensitive and specific detection of proviral bovine leukemia virus by 5' Taq nuclease PCR using a 3' minor groove binder fluorogenic probe. Journal of Virological Methods, 115 (2). pp. 167-175.

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Article Link(s): http://dx.doi.org/10.1016/j.jviromet.2003.09.029

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

Abstract

Sensitive assays are required to detect proviral bovine leukemia virus (BLV) in donor cattle used for the in vivo preparation of Australian tick fever vaccines. 5' Taq nuclease assays using 3' minor groove binder DNA probes (TaqMan®MGB) were developed and compared to conventional PCR assays for sensitive detection of Australian BLV. Seven beef and dairy herds were screened using DNA prepared by a variety of protocols to evaluate these tests. Comparative sensitivities of PCR tests were determined by testing log10 dilutions of plasmids with inserted BLV sequences. Animals were also screened by the BLV standard agar-gel immunodiffusion test (AGID) and commercial enzyme linked immunosorbent assays (ELISA) for antibodies, and an ELISA for detecting viral antigens expressed (VAE) in lymphocyte cultures. The TaqMan®MGB assay based on the pol region was the most sensitive and specific for the detection of BLV. This is the first report of a sensitive BLV 5' Taq nuclease assay.

Item Type:Article
Additional Information:© Elsevier B.V.
Keywords:Bovine leukemia virus; 5' Taq nuclease; minor groove binder; TaqMan®.
Subjects:Veterinary medicine > Veterinary virology
Veterinary medicine > Diseases of special classes of animals > Cattle
Deposited On:03 Feb 2004
Last Modified:09 Sep 2010 05:30

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