Sensitive and specific detection of bovine immunodeficiency virus and bovine syncytial virus by 5′ Taq nuclease assays with fluorescent 3′ minor groove binder-DNA probes

A.E. Lew; R.E. Bock; J. Miles; L.B. Cuttell; P. Steer; S.A. Nadin-Davis

doi:10.1016/j.jviromet.2003.10.006

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Sensitive and specific detection of bovine immunodeficiency virus and bovine syncytial virus by 5′ Taq nuclease assays with fluorescent 3′ minor groove binder-DNA probes

Journal article

Peer reviewed

Sensitive and specific detection of bovine immunodeficiency virus and bovine syncytial virus by 5′ Taq nuclease assays with fluorescent 3′ minor groove binder-DNA probes

A.E. Lew, R.E. Bock, J. Miles, L.B. Cuttell, P. Steer and S.A. Nadin-Davis

Journal of Virological Methods, Vol.116(1), pp.1-9

2004

DOI: https://doi.org/10.1016/j.jviromet.2003.10.006

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Abstract

Sensitive assays are required to detect bovine retroviruses 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 the sensitive detection of bovine syncytial virus (BSV) and bovine immunodeficiency virus (BIV). Seven beef and dairy herds were screened to evaluate these tests. Comparative sensitivities of PCR tests were determined by testing log10 dilutions of plasmids with inserts containing corresponding provirus sequences. Published PCR assays targeting BIV env sequences did not adequately amplify Australian BIV sequences. Pol sequences from Australian strains of BIV and BSV were used to design TaqMan®MGB assays, which improved sensitivity 10-fold (BIV) and 100-fold (BSV), respectively, over conventional PCR tests. This is the first report of Australian sequences of BIV and BSV and the first 5′ Taq nuclease assays described to detect these viruses. These methods could be applied to future studies requiring sensitive detection of these two bovine retroviruses.

Details

Title: Sensitive and specific detection of bovine immunodeficiency virus and bovine syncytial virus by 5′ Taq nuclease assays with fluorescent 3′ minor groove binder-DNA probes
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. Miles (Author/Creator) - Agency for Food and Fibre Sciences, Queensland Department of Primary Industries, c/o Locked Mail Bag No. 4, Moorooka 4105, Qld, Australia
L.B. Cuttell (Author/Creator) - Agency for Food and Fibre Sciences, Queensland Department of Primary Industries, c/o Locked Mail Bag No. 4, Moorooka 4105, Qld, Australia
P. Steer (Author/Creator) - Plant Industry
S.A. Nadin-Davis (Author/Creator) - Animal Diseases Research Institute
Publication Details: Journal of Virological Methods, Vol.116(1), pp.1-9
Publisher: Elsevier BV
Identifiers: 991005541410907891
Copyright: © 2003 Published by Elsevier B.V.
Murdoch Affiliation: Murdoch University
Language: English
Resource Type: Journal article

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Collaboration types: International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.66 HIV; 1.66.46 HIV Pathogenesis
Web Of Science research areas: Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology
ESI research areas: Microbiology