Abstract：
:A one-tube real-time TaqMan reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for the detection of equine arteritis virus (EAV). The test was validated using the seminal plasma and nasal secretions of infected horses that were proven to contain EAV by traditional virus isolation in rabbit kidney thirteen (RK-13) cells, as well as a variety of cell culture-propagated European and North American strains of EAV. The primers and a fluorogenic TaqMan probe were designed to amplify and detect a highly conserved region of open reading frame 7 (ORF7) of EAV. The real-time TaqMan PCR assay detected EAV RNA in all samples that were confirmed to contain infectious EAV by virus isolation. The assay had an analytical sensitivity of 10 molecules of EAV RNA allowing the detection of EAV in clinical samples or tissue culture fluid (TCF) containing at least 200 viral RNA copies per ml. Thus, the one-tube real-time TaqMan RT-PCR assay provides a rapid, accurate, quantitative, convenient and high sample throughput system for diagnosis of EAV infection, in a closed-tube format that minimizes the risk of cross-contamination.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Balasuriya UB,Leutenegger CM,Topol JB,McCollum WH,Timoney PJ,MacLachlan NJdoi
10.1016/s0166-0934(01)00416-5subject
Has Abstractpub_date
2002-03-01 00:00:00pages
21-8issue
1-2eissn
0166-0934issn
1879-0984pii
S0166093401004165journal_volume
101pub_type
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