Department of Virology, Immunobiology and Parasitology, National Veterinary Institute (SVA), Ulls väg 2B, SE-751 89 Uppsala, Sweden.
Arch Virol. 2012 May;157(5):833-44. doi: 10.1007/s00705-012-1231-0.
A novel real-time PCR strategy was applied to simultaneously detect and to discriminate low-pathogenic lentogenic and virulent meso/velogenic Newcastle disease virus (NDV). The pathotyping is achieved by a three-step semi-nested PCR. A pre-amplification of the cleavage site (CS) region of the F gene is followed by a two-level duplex real-time PCR directly targeting the CS, combining detection and pathotyping in a single tube. A wide range of NDV isolates spanning all genotypes were successfully detected and pathotyped. Clinical samples from outbreaks in Sweden in 2010 that were positive by the novel PCR method were also successfully pathotyped. The method is time-saving, reduces labour and costs and provides opportunities for rapid diagnosis at remote locations and in the field. Since the same strategy was also recently applied to avian influenza virus pathotyping, it shows promise of finding broad utility in diagnostics of infectious diseases caused by different RNA viruses in various hosts.
一种新型实时 PCR 策略被应用于同时检测和区分低致病性禽源性和强致病性中/强致病性新城疫病毒（NDV）。通过三步半巢式 PCR 实现了定型。该方法首先对 F 基因的裂解位点（CS）区域进行预扩增，然后进行二级双目标实时 PCR，在单个管中同时进行检测和定型。新型 PCR 方法成功检测和定型了涵盖所有基因型的广泛 NDV 分离株。该方法还成功对 2010 年瑞典疫情爆发的临床样本进行了定型。该方法节省时间，减少劳动力和成本，并为在偏远地区和现场进行快速诊断提供了机会。由于最近还将相同的策略应用于禽流感病毒的定型，因此它有望在不同宿主的不同 RNA 病毒引起的传染病诊断中得到广泛应用。
