Evaluation of PCR for diagnosis of bovine viral diarrhea virus in tissue homogenates.
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.
Department of Veterinary Science, University of Nebraska, Lincoln 68583.
B J
BJ
Schmitt
O J
OJ
Lopez
J F
JF
Ridpath
J
J
Galeota-Wheeler
F A
FA
Osorio
D000818:Animals; D001483:Base Sequence; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000175:diagnosis*; D002417:Cattle; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D008168:Lung / Q000382:microbiology; D008969:Molecular Sequence Data; D010576:Pestivirus / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D014420:Turbinates / Q000382:microbiology
D017931:DNA Primers; D012367:RNA, Viral