CAJ | 학술논문

本研究从2008山东表现神经症状的某新城疫疫苗免疫鸡群中分离到一株病毒.通过鸡红细胞凝集试验和血凝抑制试验,表明为新城疫病毒,将其命名为ck/CH/LSD/08-29.根据GenBank上登录的新城疫病毒基因组序列,设计14对引物经过RT-PCR扩增病毒感染的尿囊液,同时应用3'-RACE和5'-RACE技术,扩增得到该病毒全部基因组序列.结果表明,该毒株基因组由15 186个核苷酸序列组成,编码6种结构蛋白,在RNA上的顺序依次为3'-NP-P-M-F-HN-L-5'.F基因裂解位点处的氨基酸序列为~(112)R-R-Q-K-R-F~(117),符合强毒株氨基酸序列特点.通过F基因裂解位点附近高变区389个核苷酸序列构建遗传进化树,证明该毒株在分类地位上属于基因Ⅱ型.此外,病毒感染SPF鸡试验证明ck/CH/LSD/08-29毒株属于强毒嗜神经型毒株,能引起SPF鸡高致死率.
본연구종2008산동표현신경증상적모신성역역묘면역계군중분리도일주병독.통과계홍세포응집시험화혈응억제시험,표명위신성역병독,장기명명위ck/CH/LSD/08-29.근거GenBank상등록적신성역병독기인조서렬,설계14대인물경과RT-PCR확증병독감염적뇨낭액,동시응용3'-RACE화5'-RACE기술,확증득도해병독전부기인조서렬.결과표명,해독주기인조유15 186개핵감산서렬조성,편마6충결구단백,재RNA상적순서의차위3'-NP-P-M-F-HN-L-5'.F기인렬해위점처적안기산서렬위~(112)R-R-Q-K-R-F~(117),부합강독주안기산서렬특점.통과F기인렬해위점부근고변구389개핵감산서렬구건유전진화수,증명해독주재분류지위상속우기인Ⅱ형.차외,병독감염SPF계시험증명ck/CH/LSD/08-29독주속우강독기신경형독주,능인기SPF계고치사솔.
In this study,a Newcastle virus(designated as ck/CH/LSD/08-29)Was isolated from chick flock immunized with NDV vaccine which exhibited neurological symptom in Shandong province in China in 2008.Twenty of 80 day-old SPF chicks were infected with 10~(6.4) EID of the virus and all chicks died within 7 days,exhibited neurological signs.The whole genome was cloned by RT-PCR and sequenced including 3'-and 5'-terminals.The results showed that the isolate was constituted by 15 186 nt,and composed of 6 structure protein genes.The cleavage motif of F protein was~(112)R-R-Q-K-R-F~(117) which was the characteristic of velogenic strain.The evolution analysis of the 389 at sequence including the cleavage site of F gene shown that the virus was belonged to genotype Ⅱ.