Detection of bovine viral diarrhea virus by TaqMan® reverse transcription polymerase chain reaction

Carrie E. Mahlum, Sigrun Haugerud, Jan L. Shivers, Kurt D. Rossow, Sagar M. Goyal, James E. Collins, Kay S. Faaberg

Research output: Contribution to journalArticlepeer-review

62 Scopus citations

Abstract

Detection and elimination of calves and cows persistently infected with bovine viral diarrhea virus (BVDV) is important for the control of this pathogen. Historically, BVDV detection involved cell culture isolation followed by virus detection through immunofluorescence or immunoperoxidase monolayer assay (IPMA) methods. More recently, immunohistochemistry (IHC) has been added as a routine test for BVDV detection. The detection of BVDV by gel-based reverse transcription polymerase chain reaction (RT-PCR) is more sensitive and rapid than by cell culture isolation, but test results can be compromised by sample contamination during nucleic acid amplification. This study was designed to develop a closed-tube format of BVDV nucleic acid amplification and detection, TaqMan® RT-PCR. The results of this new technique were compared with those obtained with virus isolation, IPMA, and IHC. With TaqMan® RT-PCR, BVDV was detected in many samples negative by IPMA, IHC, and virus isolation with the exception of 1 sample that was positive by IHC. TaqMan® RT-PCR in a closed-tube format offers a rapid, economical, high volume, and sensitive method for BVDV detection without the concerns of amplified cDNA product contamination associated with open-tube gel-based PCR tests.

Original languageEnglish (US)
Pages (from-to)120-125
Number of pages6
JournalJournal of Veterinary Diagnostic Investigation
Volume14
Issue number2
DOIs
StatePublished - Mar 2002

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