摘要
对云南省2个发病鸡场的组织病料进行病原分离，通过血凝、血凝抑制试验和RT-PCR检测，证明分离毒株为新城疫病毒（Newcastle disease virus，NDV）。采用特异性引物经RT-PCR扩增F基因，纯化后克隆至pMD18-T载体，并对其进行测序。序列比对及系统发育分析结果表明，分离获得的云南省2株NDV毒株F基因核苷酸与La Sota株的核苷酸同源性为99．4％～99．6％，与国内外分离的流行毒株SP13和NDV027344核苷酸同源性均为99．7％～99．8％，与F48E9株的核苷酸同源性为89．0％～89．1％；F蛋白氨基酸与La Sota株的氨基酸同源性为99．3％～99．5％，与流行毒株SP13和NDV027344的氨基酸同源性均为99．5％～99．6％，与F48E9株的氨基酸同源性为91．8％～92．0％。系统发育分析结果表明，2株病毒均属于基因Ⅱ型，裂解位点氨基酸为G-R-Q-G-R L，属于弱毒株裂解位点氨基酸排列特征。
Two field strains of Newcastle disease virus （NDV） from Yunnan province had been isolated and identified bychick embryo inoculation, hemagglutination test （HA）, hemagglutination inhibition test （HI） and reverse transcriptase-poly-merase chain reaction （RT-PCR）. The F genes of two field strains were amplified by specific RT-PCR, then purified andcloned into pMD18-T vector for sequencing. The homologies of F genes of the two field strains showed 99.4% to 99.6%,99.7% to 99.8%, 89.0% to 89.1% at nucleotide level and 99.3% to 99.5%, 99.5% to 99.60%, 91.8% to 92.0% at aminoacid level with La Sota vaccine strain, SP13 and NDV027344, and F48E9 field strains, respectively. Phylogenetic analysis re-sults showed that the two strains were both belonged to genotype Ⅱ , and the amino acid sequence of cleavage site was G-R-Q-G-R-L, possessed molecular characteristics of low virulent strains of NDV.
出处
《中国畜牧兽医》
CAS
北大核心
2014年第7期54-59,共6页
China Animal Husbandry & Veterinary Medicine
基金
云南省后备人才基金(2009CI061)
云南省社会发展项目(2012CH002)
关键词
基因Ⅱ型
新城疫病毒
F基因
序列分析
genotype Ⅱ
Newcastle disease virus
F genes
sequence analysis
