Federal Research Centre for Virus Diseases of Animals, Institute of Applied Virology, Friedrich-Loeffler-Institutes, Insel Riems, Germany.
Avian Pathol. 1998;27(3):237-43. doi: 10.1080/03079459808419330.
The fusion (F) protein plays an important role in determining the virulence of Newcastle disease virus (NDV) strains. A reverse transcriptase-polymerase chain reaction (RT-PCR) is described which amplifies a 362 bp fragment encompassing the region of the F protein most important for pathogenicity. A specific PCR product was obtained independent of strain, pathogenicity and host of origin. Sequencing of the region specifying the F protein cleavage site confirmed the correlation between deduced amino acid sequence and pathogenicity. Oligonucleotides corresponding to the sequence of the pathospecific region were designed for recent German NDV isolates and labelled with digoxigenin. Hybridization of PCR fragments of different isolates with pathotype-specific oligonucleotides allowed an estimation of the pathogenicity of most isolates. Results were in good agreement with experimentally determined ICPI values.
融合(F)蛋白在决定新城疫病毒(NDV)株的毒力方面起着重要作用。本文描述了一种逆转录-聚合酶链反应(RT-PCR),可扩增包含 F 蛋白中对致病性最重要的区域的 362bp 片段。无论株系、致病性和宿主来源如何,均可获得特异性 PCR 产物。对指定 F 蛋白裂解位点的区域进行测序,证实了推导的氨基酸序列与致病性之间的相关性。针对最近德国 NDV 分离株设计了与病理特异性区域相对应的寡核苷酸,并进行了地高辛标记。不同分离株的 PCR 片段与病原体特异性寡核苷酸的杂交可估计大多数分离株的致病性。结果与实验确定的 ICPI 值非常吻合。