伪狂犬病病毒Guizhou-T1株的分离鉴定及其遗传变异研究
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摘要
为了解贵州猪伪狂犬病病毒(PRV)流行株的病原形态特征、致病性及遗传变异情况,本实验对收集自贵州规模化猪场伪狂犬病净化过程中PCR检测的阳性病料样品,采用细胞接毒分离培养、病毒的形态结构观察和易感动物接种试验对其病原进行鉴定。结果显示,本研究分离到一株PRV,命名为PRV Guizhou-T1株;并对其进行病毒效价的测定及g E基因核苷酸和氨基酸序列差异性分析。结果显示:PRV Guizhou-T1株gE基因序列与GZ-Z1株和Guizhou-DY株同源性分别为97.5%和99.3%,其TCID_(50)为10~(-10.11)/100μL;PRV Guizhou-T1株在gE基因编码的氨基酸序列第48和第496位均有天冬氨酸(Asp D)的插入,与PRV变异株变异位点一致,且在其第48位缺失酪氨酸(Tyr Y)。表明本研究分离得到了一株PRV变异株。本研究可为贵州PRV病原学的研究提供参考。
A pseudorabies virus(PRV) was isolated from the positive sample collected from a large scale pig farm in Guizhou and identified by PCR and sequence analysis. The typical PRVs were observed in infected cell by electron microscope. The alignment of the gE gene sequence showed that the isolate was high similarity(97.7%-99.8%) with other PRV reference strains available in GenBank. The virus titer was 10~(-10.11) TCID_(50) in Vero cells. The virus isolation could be useful for further study of epidemiology and molecular biology of PRV.
A pseudorabies virus(PRV) was isolated from the positive sample collected from a large scale pig farm in Guizhou and identified by PCR and sequence analysis. The typical PRVs were observed in infected cell by electron microscope. The alignment of the gE gene sequence showed that the isolate was high similarity(97.7%-99.8%) with other PRV reference strains available in GenBank. The virus titer was 10~(-10.11) TCID_(50) in Vero cells. The virus isolation could be useful for further study of epidemiology and molecular biology of PRV.
引文
[1]郝飞,汤德元,李春燕,等.伪狂犬病病毒Guizhou-DY株的分离鉴定及致病性研究[J].畜牧兽医学报,2013,44(11):1851-1856.
[2]郝飞,张华,汤德元,等.我国规模化猪场主要病毒性疫病的综合防控对策[J].畜牧与兽医,2012,44(10):86-89.
[3]Boeleart F,Deluyker H,Maes D.Prevalence of herds with young sows seropositive to pseudorabies(Aujeszkyps disease)in northern Belgium[J].Prev Vet Med,1999,41(4):239-255.
[4]祖立闯,谢金文,王文秀,等.我国猪伪狂犬病的流行情况分析[J].养猪,2017,41(03):123-128.
[5]Tamba M,Calabrese R,F inell I E.Risk factors for Aujeszkyps disease seropositivity of swine herds of a region of northern Italy[J].Prev Vet Med,2002,54(3):203-212.
[6]李达,马萍,汤德元,等.规模化猪场猪瘟和伪狂大病的净化方案[J].畜牧与兽医,2015,47(7):119-123.
[7]王继春,曾容愚,Daniel T,等.猪伪狂犬病活疫苗(Bartha K61株)对变异株的保护效力[J].畜牧与兽医,2015,47(12):1-4.
[8]曾智勇,周莉,汤德元,等.猪伪狂犬病病毒GZ-Z1株的分离鉴定[J].中国兽医科学,2011,41(5):459-463.
[9]彭金美,安同庆,赵洪远,等.猪伪狂犬病病毒新流行株的分离鉴定及抗原差异性分析[J].中国预防兽医学报,2013,35(01):1-4.
[10]赵鸿远,彭金美,安同庆,等.猪伪狂犬病病毒变异株的分离鉴定及其gE基因的分子特征[J].中国预防兽医学报,2014,36(7):506-509.
[11]张青占.变异猪伪狂犬病毒的分离鉴定及生物学特性分析[D].中国农业科学院,2013.
[12]Yu Xiu-ling,Zhou Zhi,Hu Dong-mei,et al.Pathogenic Pseudorabies Virus,China,2012[J].Emerg Infect Dis,2014,20(08):102-104.
[13]钟承,刘蕾,张乐天,等.伪狂犬病病毒BJ/YT株毒力相关基因gE和TK序列分析[J].中国畜牧兽医,2014,41(11):73-79.
[2]郝飞,张华,汤德元,等.我国规模化猪场主要病毒性疫病的综合防控对策[J].畜牧与兽医,2012,44(10):86-89.
[3]Boeleart F,Deluyker H,Maes D.Prevalence of herds with young sows seropositive to pseudorabies(Aujeszkyps disease)in northern Belgium[J].Prev Vet Med,1999,41(4):239-255.
[4]祖立闯,谢金文,王文秀,等.我国猪伪狂犬病的流行情况分析[J].养猪,2017,41(03):123-128.
[5]Tamba M,Calabrese R,F inell I E.Risk factors for Aujeszkyps disease seropositivity of swine herds of a region of northern Italy[J].Prev Vet Med,2002,54(3):203-212.
[6]李达,马萍,汤德元,等.规模化猪场猪瘟和伪狂大病的净化方案[J].畜牧与兽医,2015,47(7):119-123.
[7]王继春,曾容愚,Daniel T,等.猪伪狂犬病活疫苗(Bartha K61株)对变异株的保护效力[J].畜牧与兽医,2015,47(12):1-4.
[8]曾智勇,周莉,汤德元,等.猪伪狂犬病病毒GZ-Z1株的分离鉴定[J].中国兽医科学,2011,41(5):459-463.
[9]彭金美,安同庆,赵洪远,等.猪伪狂犬病病毒新流行株的分离鉴定及抗原差异性分析[J].中国预防兽医学报,2013,35(01):1-4.
[10]赵鸿远,彭金美,安同庆,等.猪伪狂犬病病毒变异株的分离鉴定及其gE基因的分子特征[J].中国预防兽医学报,2014,36(7):506-509.
[11]张青占.变异猪伪狂犬病毒的分离鉴定及生物学特性分析[D].中国农业科学院,2013.
[12]Yu Xiu-ling,Zhou Zhi,Hu Dong-mei,et al.Pathogenic Pseudorabies Virus,China,2012[J].Emerg Infect Dis,2014,20(08):102-104.
[13]钟承,刘蕾,张乐天,等.伪狂犬病病毒BJ/YT株毒力相关基因gE和TK序列分析[J].中国畜牧兽医,2014,41(11):73-79.