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United States Department of Agriculture

Agricultural Research Service

Title: Nested Reverse Transcriptase-Polymerase Chain Reaction (Rt-Pcr) for Typing Ruminant Pestiviruses: Bovine Viral Diarrhea Viruses and Border Disease Virus

Authors
item Fulton, R - OKLAHOMA STATE UNIV
item D'Offay, J - OKLAHOMA STATE UNIV
item Saliki, J - OKLAHOMA STATE UNIV
item Burge, L - OKLAHOMA STATE UNIV
item Helman, R - OKLAHOMA STATE UNIV
item Confer, A - OKLAHOMA STATE UNIV
item Bolin, Steven
item Ridpath, Julia

Submitted to: Canadian Journal of Veterinary Research
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: August 25, 1999
Publication Date: N/A

Interpretive Summary: Bovine viral diarrhea virus (BVDV) is a widespread viral pathogen of cattle that induces respiratory, reproductive, and enteric disease. Diagnosis of BVDV infections of cattle is based on observation of clinical signs of disease and on laboratory detection of viral antigen or viral nucleic acid in clinical specimens from diseased cattle. A rapid nucleic acid-based diagnostic test, using polymerase chain reaction amplification (PCR) of viral RNA, was evaluated against 30 strains of virus. The PCR test was designed to detect RNA from BVDV. Additionally, the test can detect RNA from a closely related virus of sheep, border disease virus (BDV). To enhance the usefulness of the test in disease outbreaks, a second round of PCR was included in the test so that when the test was concluded the taxonomic viral group to which the BVDV or BDV belonged was identified. The PCR test accurately detected each virus and correctly identified the taxonomic group to which each virus had been assigned by previous analyses. This test should be useful to veterinary diagnostic laboratories for detection and classification of BVDV and BDV. Also, the test should be useful to veterinary biologics manufacturers as a quality control measure to detect adventitious BVDV or BDV in veterinary vaccines. Rapid and accurate diagnostic tests benefit cattle producers by helping to identify disease-producing agents so that proper control measures can be used in the herd.

Technical Abstract: The Pestivirus genus of the family Flaviviridae contains the viruses causing illnesses in the U.S. livestock operations: bovine viral diarrhea virus (BVDV) types 1 and 2 and border disease virus (BDV). Although serologic differences may be evident, there is also serologic cross reactivity among these three ruminant pestiviruses. A nested reverse transcription-polymerase chain reaction (RT-PCR) assay was used to distinguish reference strains of BVDV types 1 and 2 and BDV. The RT-PCR detected BVDV types 1 and 2 among a select number of viruses from diagnostic laboratory accessions. Also, the RT-PCR identified the type of BVDV present in multivalent modified live virus (MLV) bovine viral vaccines containing four separate viruses including BVDV. This RT-PCR assay is useful to detect the respective ruminant pestivirus types present in reference strains, diagnostic laboratory accessions, and MLV vaccines.

Last Modified: 8/30/2014
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