Development of a duplex quantitative polymerase chain reaction assay for detection of bovine herpesvirus 1 and bovine viral diarrhea virus in bovine follicular fluid.
The objective of this study was to develop a duplex quantitative polymerase chain reaction (qPCR) assay for simultaneous detection of bovine herpesvirus 1 (BoHV-1) and bovine viral diarrhea virus (BVDV) type I and type II. Follicular fluid was collected from a BoHV-1 acutely infected heifer, a BVDV I persistently infected heifer, and from 10 ovaries recovered from an abattoir. Both the BoHV-1 and BVDV contaminated follicular fluid were diluted 1:5 to 1:10(7) using the pooled, abattoir-origin follicular fluid. Each dilution sample was analyzed using the duplex qPCR, virus isolation, reverse transcription-nested PCR (RT-nPCR), and BoHV-1 qPCR. The duplex qPCR was able to simultaneously detect BoHV-1 and BVDV I in the fluid diluted to 1:100 and 1:1000, respectively. These results corresponded with the reverse transcription-nested PCR and BoHV-1 qPCR. Therefore, the duplex qPCR might be used for quality assurance testing to identify these two viruses in cells, fluids and tissues collected from donor animals and used in reproductive technologies.
Marley
Mylissa S D
MS
Department of Pathobiology, College of Veterinary Medicine, 127 Sugg Laboratory, Auburn University, Auburn, AL 36849, USA.
Givens
M Daniel
MD
Galik
Patricia K
PK
Riddell
Kay P
KP
Stringfellow
David A
DA
D000818:Animals; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000821:virology; D002417:Cattle; D001908:Diarrhea Viruses, Bovine Viral / Q000302:isolation & purification*; D005260:Female; D015571:Follicular Fluid / Q000821:virology*; D007242:Herpesvirus 1, Bovine / Q000302:isolation & purification*; D007241:Infectious Bovine Rhinotracheitis / Q000821:virology; D016133:Polymerase Chain Reaction / Q000662:veterinary*