Development of a real time RT-PCR to detect and type ovine pestiviruses.
A real time one-step RT-PCR was designed to detect and type border disease virus (BDV), bovine viral diarrhea virus (BVDV) type 1 and BVDV type 2 in ovine samples. The real time RT-PCR was shown to behave in a linear manner and had limits of detection of 100-1000 copies of viral RNA as judged by in vitro transcribed RNA. The real time RT-PCR was validated on 50 clinical samples from UK flocks and was more sensitive than a virus isolation and a classical nested RT-PCR (nRT-PCR). The results of real time RT-PCR virus typing agreed completely with sequencing. The majority of ovine isolates were BDV; a small proportion were BVDV type 1. BVDV type 2 was not detected in any sample. This test appears reliable and can be used for the typing of ovine pestiviruses in the UK.
Willoughby
K
K
Virus Surveillance Unit, Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ, UK. willk@mri.sari.ac.uk
Valdazo-González
B
B
Maley
M
M
Gilray
J
J
Nettleton
P F
PF
D000818:Animals; D001882:Border Disease / Q000175:diagnosis* / Q000821:virology; D018083:Border disease virus / Q000145:classification / Q000302:isolation & purification*; D030241:Diarrhea Virus 1, Bovine Viral / Q000145:classification / Q000302:isolation & purification*; D030242:Diarrhea Virus 2, Bovine Viral / Q000145:classification / Q000302:isolation & purification*; D018182:Pestivirus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D010802:Phylogeny; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis / Q000821:virology; D006113:United Kingdom
D012367:RNA, Viral