Willoughby K, Valdazo-González B, Maley M, Gilray J, Nettleton P F
Virus Surveillance Unit, Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Midlothian EH26 0PZ, UK.
J Virol Methods. 2006 Mar;132(1-2):187-94. doi: 10.1016/j.jviromet.2005.10.007. Epub 2005 Nov 23.
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.
设计了一种实时一步法逆转录聚合酶链反应（RT-PCR），用于检测绵羊样本中的边界病病毒（BDV）、1型牛病毒性腹泻病毒（BVDV）和2型BVDV，并对其进行分型。实时RT-PCR呈线性反应，通过体外转录RNA判断，其检测限为100 - 1000个病毒RNA拷贝。该实时RT-PCR在来自英国羊群的50份临床样本上得到验证，比病毒分离和经典巢式RT-PCR（nRT-PCR）更灵敏。实时RT-PCR病毒分型结果与测序结果完全一致。大多数绵羊分离株为BDV；一小部分为1型BVDV。在任何样本中均未检测到2型BVDV。该检测方法似乎可靠，可用于英国绵羊瘟病毒的分型。
