摘要
为了确定引起绵羊呼吸道传染病的病原,本研究于2019年对一患急性呼吸道疾病死亡绵羊的病料进行了病毒分离,从其胸腔积液中分离到一株绵羊副流感病毒3型(OPIV3),命名为PE2019株。该病毒能够在MDBK细胞上增殖,产生细胞圆缩、集聚成簇等为特征的细胞病变;电子显微镜观察可见OPIV3 PE2019株为直径约300 nm的圆形粒子;经检测该分离株病毒滴度为107.8TCID50/mL。分离株对小鼠和豚鼠的红细胞血凝价均为164,不凝集鸡、兔、绵羊和牛的红细胞。参考山羊副流感病毒3型全基因组序列(MK091103),设计其N和M基因特异性引物,通过RT-PCR可从病毒细胞培养物中扩增到特异性片段,对其进行序列分析,结果显示,分离株N基因序列与山羊、牛和人副流感病毒3型参考株的基因同源性分别为84.0%、82.1%和78.8%,M基因序列与山羊、牛和人副流感病毒3型参考株的基因同源性分别为81.5%、78.4%和77.6%,OPIV3 N和M基因均为独立一个分支。本研究首次于国内分离到OPIV3,为了解国内OPIV3的流行情况及疫苗的研制奠定了基础。
In order to identify the pathogens of respiratory infectious disease in sheep,viruses were isolated from the samples of a dead sheep with acute respiratory disease clinical signs in 2019.A strain of ovine parainfluenza virus type 3(OPIV3)was isolated from the pleural effusion and named PE2019.This strain could proliferate in the MDBK cells and developed cell shrinkage,clustering and cell lesions.Under the electron microscope,small round particles with a diameter of about 300 nm could be observed.The titer of this isolated strain was 107.8 TCID50/mL.The hemagglutination titer of this strain to the red blood cells of mice and guinea pigs were 164,and the hemagglutination to the red blood cells of chicken,rabbit,sheep and cattle was not observed.The specific primersfor N and M genes were designed with reference to the sequences of Caprine parainfluenza virus 3 strain GS2017-2(MK091103)genome,and specific fragments for the N and M genes were amplified by RT-PCR for sequence analysis.Results showed that NA gene homology of this isolate with the reference strains of CPIV3、BPIV3 and HPIV3 were 84.0%,82.1%and 18.8%,respectively,and M genehomology of the virus with the three reference strains were 81.5%,78.4%and 77.6%,respectively.The N and M genes of the isolated virus formed separate branches in the phylogenetic trees,respectively.Taken together,our study firstly isolated a strain of OPIV3 in sheep in China and lays the foundation for further study on the epidemics of OPIV3 and vaccine development.
作者
朱远茂
陈瑞红
林俊
杨木娇
薛飞
ZHU Yuan-mao;CHEN Rui-hong;LIN Jun;YANG Mu-jiao;XUE Fei(State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,ChineseAcademy ofAgricultural Sciences,Harbin 150069,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2020年第5期434-438,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
国家重点研发计划课题:牛羊虫媒病和慢病毒病的诊断与检测新技术研究(2016YFD0500908)
国家自然科学基金(31372452)
关键词
绵羊副流感病毒3型
分离
鉴定
ovien parainfluenza virus type 3
isolation
identification
