Evaluation of PCR for diagnosis of bovine viral diarrhea virus in tissue homogenates

Authors

Beverly Schmitt, Department of Veterinary Science, University of Nebraska
Osvaldo Lopez, Department of Veterinary Science, University of Nebraska
Julia Ridpath, Virology Cattle Research Unit, National Animal Disease Center, Agricultural Research Service, USDA
Judith Galeota-Wheeler, Department of Veterinary Science, University of Nebraska
Fernando A. Osorio, Department of Veterinary Science, University of NebraskaFollow

Date of this Version

1994

Citation

Journal of Veterinary Diagnostic Investigation, January 1994; vol. 6, 1: pp. 44-47.

Comments

This article is a U.S. government work, and is not subject to copyright in the United States.

Abstract

Tissue homogenates from 60 specimens submitted to the Veterinary Diagnostic Center were evaluated by polymerase chain reaction (PCR) for detection of bovine viral diarrhea virus (BVDV). Conventional virus isolation procedures showed the specimens contained BVDV. The BVDV RNA was extracted from the homogenates and subjected to a reverse transcription reaction followed by PCR amplification. The PCR product was blotted onto a nylon membrane and hybridized with a 30-base pair oligonucleotide probe labeled with 32P. One set of PCR primers detected BVDV in 46/60 (77%) of the tissue homogenates. An additional set of primers was used to detect 10/11 samples that had escaped detection with the first set of primers. The results indicate that BVDV can be detected by PCR directly out of tissue homogenates generated in a diagnostic setting.