中国畜牧兽医

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云南2株基因Ⅱ型新城疫病毒F基因变异特性分析

张文东1,宋建领2,赵焕云1,刘庆亮3,胡媛媛4,曾伟3,张富强5   

  1. (1.云南省动物疫病预防控制中心,云南昆明  6500512.云南省热带亚热带动物病毒病重点实验室,云南昆明  650224 3.云南农业大学, 云南昆明  6502234.云南农业职业技术学院,云南昆明  6502205.成都军区疾病预防控制中心, 云南昆明  650032)

  • 收稿日期:2014-01-09 出版日期:2014-07-20 发布日期:2014-08-21
  • 通讯作者: 宋建领。E-mail:jianling_song@hotmail.com;Tel:0871-65016575 张富强。E-mail:zfq1968@yahoo.com.cn;Tel:0871-64775708
  • 作者简介:张文东(1973—),男,云南人,硕士,高级兽医师,研究方向:动物疾病诊断。
  • 基金资助:

    云南省后备人才基金(2009CI061);云南省社会发展项目(2012CH002)。

Mutation Characteristics Analysis of F Genes of Two Strains Genotype Ⅱ Newcastle Disease Virus from Yunnan Province

ZHANG Wen-dong1,SONG Jian-ling2,ZHAO Huan-yun1,LIU Qing-liang3,HU Yuan-yuan4,ZENG Wei3, ZHANG Fu-qiang5   

  1. (1.Center for Animal Disease Prevention and Control in Yunnan Province, Kunming 650051, China; 2.Yunnan Tropical and Subtropical Animal Virus Diseases Laboratory, Kunming 650224, China; 3.Yunnan Agricultural University, Kunming 650223, China; 4.Yunnan Vocational and Technical College of Agriculture, Kunming 650220, China5.Center for Disease Prevention and Control, Chengdu Military Region, Kunming 650032, China)

  • Received:2014-01-09 Online:2014-07-20 Published:2014-08-21

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。

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Abstract: Two field strains of Newcastle disease virus(NDV) from Yunnan province had been isolated and identified by chick embryo inoculation, hemagglutination test(HA), hemagglutination inhibition test(HI) and reverse transcriptase-polymerase chain reaction(RT-PCR). The F genes of two field strains were amplified by specific RT-PCR, then purified and cloned 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 99.6%, 91.8% to 92.0% at amino acid level with La Sota vaccine strain, SP13 and NDV027344, and F48E9 field strains, respectively. Phylogenetic analysis results showed that the two strains were both belonged to genotypeⅡ, and the amino acid sequence of cleavage site was G-R-Q-G-R-L, possessed molecular characteristics of low virulent strains of NDV.

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