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

A.E. Lew; R.E. Bock; J.B. Molloy; C.M. Minchin; S.J. Robinson; P. Steer

doi:10.1016/j.jviromet.2003.09.029

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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

Journal article

Peer reviewed

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

A.E. Lew, R.E. Bock, J.B. Molloy, C.M. Minchin, S.J. Robinson and P. Steer

Journal of Virological Methods, Vol.115(2), pp.167-175

2004

DOI: https://doi.org/10.1016/j.jviromet.2003.09.029

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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.

Details

Title: Sensitive and specific detection of proviral bovine leukemia virus by 5′ Taq nuclease PCR using a 3′ minor groove binder fluorogenic probe
Authors/Creators: A.E. Lew (Author/Creator) - Agency for Food and Fibre Sciences, Queensland Department of Primary Industries, c/o Locked Mail Bag No. 4, Moorooka 4105, QLD, Australia
R.E. Bock (Author/Creator) - Plant Industry
J.B. Molloy (Author/Creator) - Agency for Food and Fibre Sciences, Queensland Department of Primary Industries, c/o Locked Mail Bag No. 4, Moorooka 4105, QLD, Australia
C.M. Minchin (Author/Creator) - Agency for Food and Fibre Sciences, Queensland Department of Primary Industries, c/o Locked Mail Bag No. 4, Moorooka 4105, QLD, Australia
S.J. Robinson (Author/Creator) - Plant Industry
P. Steer (Author/Creator) - Plant Industry
Publication Details: Journal of Virological Methods, Vol.115(2), pp.167-175
Publisher: Elsevier BV
Identifiers: 991005541547407891
Copyright: © 2015 Elsevier B.V.
Murdoch Affiliation: Murdoch University
Language: English
Resource Type: Journal article

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Citation topics: 1 Clinical & Life Sciences; 1.66 HIV; 1.66.1464 HTLV-1/BLV Pathogenesis
Web Of Science research areas: Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology
ESI research areas: Microbiology