Detection of bovine viral diarrhea virus (BVDV) in single or small groups of preimplantation bovine embryos.
The objectives of this study were to develop techniques to detect BVDV associated with single or small groups of bovine embryos contained in small aliquots of medium using either virus isolation (VI) or real time quantitative polymerase chain reaction (RT-QPCR) assays. In vivo-derived and in vitro-produced bovine embryos at 7 d post-fertilization were exposed to SD-1, a high affinity strain of BVDV, for 2 h and then processed according to the International Embryo Transfer Society (IETS) guidelines prior to testing. Groups of five or two in vivo-derived embryos, and single in vivo-derived embryos, were VI positive for BVDV 100, 50, and 33% of the time, and were RT-QPCR positive 100, 75, and 42% of the time, respectively. The virus was detected by the VI technique in all of the groups of five or two in vitro-produced embryos and in all of the single in vitro-produced embryos, and it was detected in 100, 80, and 50%, using RT-QPCR. Techniques for RT-QPCR were sufficiently sensitive to detect 10 copies of viral RNA in a sample and to detect BVDV associated with single embryos. Application of this new technology, RT-QPCR, will facilitate additional studies to further assess the risk of transmission of BVDV through embryo transfer.
Gard
J A
JA
Department of Clinical Sciences, College of Veterinary Medicine, Auburn University, AL 36849, USA. waldrja@vetmed.auburn.edu
Givens
M D
MD
Riddell
K P
KP
Galik
P K
PK
Zhang
Y
Y
Stringfellow
D A
DA
Marley
M S D
MS
D000818:Animals; D001755:Blastocyst / Q000821:virology*; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000517:prevention & control / Q000635:transmission; D002417:Cattle / Q000196:embryology* / Q000821:virology*; D046508:Culture Techniques; D001908:Diarrhea Viruses, Bovine Viral / Q000302:isolation & purification*; D005260:Female; D005307:Fertilization in Vitro; D018445:Infectious Disease Transmission, Vertical / Q000662:veterinary; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity