title: Development of a duplex quantitative polymerase chain reaction assay for detection of bovine herpesvirus 1 and bovine viral diarrhea virus in bovine follicular fluid.
abstract: 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.
authors.affiliation: Department of Pathobiology, College of Veterinary Medicine, 127 Sugg Laboratory, Auburn University, Auburn, AL 36849, USA.
authors.forename: Mylissa S D
authors.initials: MS
authors.lastname: Marley
authors.forename: M Daniel
authors.initials: MD
authors.lastname: Givens
authors.forename: Patricia K
authors.initials: PK
authors.lastname: Galik
authors.forename: Kay P
authors.initials: KP
authors.lastname: Riddell
authors.forename: David A
authors.initials: DA
authors.lastname: Stringfellow
mesh_terms: 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*