猪瘟病毒GZA株的分离鉴定及遗传变异分析
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摘要
本研究收集贵州规模化猪场猪瘟净化过程中RT-PCR检测阳性病料,采用细胞接毒技术、病毒的形态结构观察和间接免疫荧光试验等分离鉴定了1株猪瘟病毒(CSFV),命名为CSFV GZA株,并对GZA株E2基因的核苷酸和氨基酸序列进行差异性分析。分离鉴定结果表明:接种PK-15细胞后连传7代RT-PCR均能扩增出CSFV特异性条带;病毒粒子在电镜下呈椭圆形或圆形,直径30～80nm;该毒株E2基因与参考毒株E2基因的核苷酸同源性82.8%～83.4%,氨基酸相似度88.7%～90.6%,并在713,725,729,734,738氨基酸位点发生改变;遗传进化树分析还得出GZA株E2基因与参考毒株遗传距离较远。说明本次分离株与其他流行株存在一定的差异,可为以后CSFV病原学的研究提供参考。
A classical swine fever virus was isolated from the positive sample of large scale pig farm in Guizhou and identified by RT-PCR and sequence analysis.The classical swine fever virus were observed in infected cell by electron microscope and the E2 gene sequence was analyzed.The alignment of the E2 sequence showed that the isolate shared high similarity(82.8%-83.4%)with other CSFV reference strains available in GenBank.These results could be useful for the research of epidemiology and molecular biology of CSFV.
A classical swine fever virus was isolated from the positive sample of large scale pig farm in Guizhou and identified by RT-PCR and sequence analysis.The classical swine fever virus were observed in infected cell by electron microscope and the E2 gene sequence was analyzed.The alignment of the E2 sequence showed that the isolate shared high similarity(82.8%-83.4%)with other CSFV reference strains available in GenBank.These results could be useful for the research of epidemiology and molecular biology of CSFV.
引文
[1]蔡宝祥.猪瘟的免疫综述[J].动物科学与动物医学,2005,22(2):24-27.
[2]殷震,刘景华.动物病毒学[M].2版.北京:科学出版社,1997:652-664.
[3]万强.当前动物疫病防治形势与对策-《国家中长期动物疫病防治规划(2012-2020年)》学习体会[J].中国动物检疫,2013,30(8):1-3,13.
[4]李达,马萍,汤德元,等.规模化猪场猪瘟和伪狂犬病的净化方案[J].畜牧与兽医,2015,47(7):119-123.
[5]POSTEL A,SCHMEISER S,BERNAU J,et al.Improved strategy for phylogenetic analysis of classical swine fever virus based on full-length E2encoding sequences[J].Vet Res,2012,43(1):50.
[6]谭颖翌,毛燕,吴健敏,等.1986-2011年广西猪瘟流行监测及遗传变异分析[J].中国兽医学报,2013,33(10):1481-1487.
[7]JIANG D L,GONG W,LI R C,et al.Phylogcnctic analysis using E2gene of classical swine fever virus revcals a new subgenotype in China[J].Infect Genet Evol,2013,17(2):231-238.
[8]彭志成,龚文杰,吕宗吉,等.广东省猪瘟病毒流行毒株遗传多样性分析[J].中国兽医学报,2014,34(6):894-903.
[9]冯丽苹.2014-2015年我国猪瘟病毒的分子流行病学分析[D].湖北荆州:长江大学,2016.
[10]陈田姣,何奇松,颜健华,等.广西猪瘟病毒E0和E2基因的克隆及序列分析[J].中国畜牧兽医,2015,42(4):789-795.
[11]吴军,袁小宁,杨可妍,等.广西地区猪瘟流行毒株E2基因的序列测定与分析[J].南方农业学报,2014,45(11):2065-2070.
[12]张瑞琴.山西猪瘟抗体水平调查及野毒株E2遗传变异多样性分析[D].吉林长春:吉林大学,2014.
[13]姚敬明,孟帆,雷宇平,等.山西猪瘟野毒株E2基因序列测定与遗传变异多样性分析[J].中国兽医学报,2014,34(9):1405-1410.
[14]魏春华,刘建奎,戴爱玲,等.福建地区猪瘟病毒流行株基因分型及E2基因遗传变异分析[J].中国兽医学报,2016,36(12):2009-2013.
[15]李栋梁,王新港,郭天准,等.2014-2015年河南省猪瘟野毒E2、E0基因变异分析[J].中国兽医学报,2017,37(7):1212-1219.
[2]殷震,刘景华.动物病毒学[M].2版.北京:科学出版社,1997:652-664.
[3]万强.当前动物疫病防治形势与对策-《国家中长期动物疫病防治规划(2012-2020年)》学习体会[J].中国动物检疫,2013,30(8):1-3,13.
[4]李达,马萍,汤德元,等.规模化猪场猪瘟和伪狂犬病的净化方案[J].畜牧与兽医,2015,47(7):119-123.
[5]POSTEL A,SCHMEISER S,BERNAU J,et al.Improved strategy for phylogenetic analysis of classical swine fever virus based on full-length E2encoding sequences[J].Vet Res,2012,43(1):50.
[6]谭颖翌,毛燕,吴健敏,等.1986-2011年广西猪瘟流行监测及遗传变异分析[J].中国兽医学报,2013,33(10):1481-1487.
[7]JIANG D L,GONG W,LI R C,et al.Phylogcnctic analysis using E2gene of classical swine fever virus revcals a new subgenotype in China[J].Infect Genet Evol,2013,17(2):231-238.
[8]彭志成,龚文杰,吕宗吉,等.广东省猪瘟病毒流行毒株遗传多样性分析[J].中国兽医学报,2014,34(6):894-903.
[9]冯丽苹.2014-2015年我国猪瘟病毒的分子流行病学分析[D].湖北荆州:长江大学,2016.
[10]陈田姣,何奇松,颜健华,等.广西猪瘟病毒E0和E2基因的克隆及序列分析[J].中国畜牧兽医,2015,42(4):789-795.
[11]吴军,袁小宁,杨可妍,等.广西地区猪瘟流行毒株E2基因的序列测定与分析[J].南方农业学报,2014,45(11):2065-2070.
[12]张瑞琴.山西猪瘟抗体水平调查及野毒株E2遗传变异多样性分析[D].吉林长春:吉林大学,2014.
[13]姚敬明,孟帆,雷宇平,等.山西猪瘟野毒株E2基因序列测定与遗传变异多样性分析[J].中国兽医学报,2014,34(9):1405-1410.
[14]魏春华,刘建奎,戴爱玲,等.福建地区猪瘟病毒流行株基因分型及E2基因遗传变异分析[J].中国兽医学报,2016,36(12):2009-2013.
[15]李栋梁,王新港,郭天准,等.2014-2015年河南省猪瘟野毒E2、E0基因变异分析[J].中国兽医学报,2017,37(7):1212-1219.