河北北部猪圆环病毒2型流行毒株的遗传变异分析
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摘要
为了掌握河北北部地区猪圆环病毒2型(PCV2)流行病学特点和遗传变异情况,应用PCR检测方法对2015—2017年间收集的河北北部猪场86份临床病料进行PCV2检测,并对阳性毒株的全基因进行扩增、克隆和测序。结果显示,86份临床样品中,有38份样品为PCV2阳性,阳性率为44.19%,选取11株进行病毒全基因扩增,并利用序列比对软件进行遗传变异分析发现,11株病毒之间同源性为95.9%～99.9%,与各参考毒株之间同源性为93.5%～99.9%。在遗传进化关系上,11株病毒分别处于2个分支:PCV2b和PCV2d,表明河北北部地区PCV2流行比较广泛,并且以PCV2b和PCV2d毒株流行为主。本研究充实了河北北部地区PCV2流行病学调查资料,也为河北PCV2疫苗株的研发和选择提供了依据。
In order to investigate molecular epidemiology and genetic variation of porcine circovirus type 2(PCV2),a total of 86 clinical samples were collected between 2015 and 2017 in northern Hebei province.All these clinical samples were subjected to PCR,and the positive samples were amplified and cloned,then sequenced to obtain the whole genome of PCV2.The results demonstrated that the positive rate of PCV2 was 44.19%(38/86).These 11 strains of PCV2-positive samples were selected to sequence the complete genome,and to process genetic variation analysis using sequence alignment software.Sequence comparison showed that the nucleotide sequence homologies among 11 PCV2 strains were up to 95.9%-99.9%,and were up to 93.5%-99.9%compared with the reference strains.The phylogenetic analysis revealed that these 11 strains belonged to two subgenotypes:PCV2 band PCV2 d,indicating that PCV2 was endemic in northern Hebei province with dominant strains of PCV2 band PCV2 d.This study significantly enriched the PCV2 epidemiological survey in northern Hebei province,and provided the basis for the development and selection of PCV2 vaccines.
In order to investigate molecular epidemiology and genetic variation of porcine circovirus type 2(PCV2),a total of 86 clinical samples were collected between 2015 and 2017 in northern Hebei province.All these clinical samples were subjected to PCR,and the positive samples were amplified and cloned,then sequenced to obtain the whole genome of PCV2.The results demonstrated that the positive rate of PCV2 was 44.19%(38/86).These 11 strains of PCV2-positive samples were selected to sequence the complete genome,and to process genetic variation analysis using sequence alignment software.Sequence comparison showed that the nucleotide sequence homologies among 11 PCV2 strains were up to 95.9%-99.9%,and were up to 93.5%-99.9%compared with the reference strains.The phylogenetic analysis revealed that these 11 strains belonged to two subgenotypes:PCV2 band PCV2 d,indicating that PCV2 was endemic in northern Hebei province with dominant strains of PCV2 band PCV2 d.This study significantly enriched the PCV2 epidemiological survey in northern Hebei province,and provided the basis for the development and selection of PCV2 vaccines.
引文
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[2]FENAUX M,HALBUR P G,GILL M,et al.Genetic characterization of type 2porcine circovirus(PCV-2)from pigs with postweaning multisystemic wasting syndrome in different geographic regions of North America and development of a differential PCR-restriction fragment length polymorphism assay to detect and differentiate between infections with PCV-1and PCV-2[J].J Clin Microbiol,2000,38(7):2494-2503.
[3]HE J L,CAO J J,ZHOU N,et al.Identification and functional analysis of the novel ORF4protein encoded by porcine circovirus type 2[J].J Virol,2013,87(3):1420-1429.
[4]DUPONT K,NIELSEN E O,BAEKBO P,et al.Genomic analysis of PCV2isolates from Danish archives and a current PMWS case-control study supports a shift in genotypes with time[J].Vet Microbiol,2008,128(1/2):56-64.
[5]OLVERA A,CORTEY M,SEGALES J.Molecular evolution of porcine circovirus type 2genomes:phylogeny and clonality[J].Virology,2007,357(2):175-185.
[6]陈光达,许信刚,童德文.PCV-2、PPV和PRV多重PCR检测方法的建立及初步应用[J].西北农林科技大学学报(自然科学版),2011,39(11):1-6.
[7]CHEUNG A K,BOLIN S R.Kinetics of porcine circovirus type 2replication[J].Arch Virol,2002,147(1):43-58.
[8]江戈龙,高文超,刘德武,等.母源抗体对感染PCV2仔猪体内病毒变化和免疫反应的影响[J].中国兽医学报,2017,37(3):398-403.
[9]李然,赵紫印,潘广林,等.1只秦岭地区野外捕获野猪重要猪源病毒的检测及PCV2分离株基因序列分析[J].中国兽医学报,2017,37(10):1835-1840.
[10] GE X N,WANG F,GUO X,et al.Porcine circovirus type 2and its associated diseases in China[J].Virus Res,2012,164(1/2):100-106.
[11] JIANG C G,WANG G,TU Y B,et al.Genetic analysis of porcine circovirus type 2in China[J].Arch Virol,2017,162(9):2715-2726.
[12] LI L M,YUAN W Z,GUO H Y,et al.Prevalence and genetic variation of porcine circovirus type 2in Hebei,China from 2004to 2014[J].Gene,2016,586(2):222-227.
[13]吴忻,孟帆,姚敬明,等.猪圆环病毒2型山西分离株的遗传变异多样性分析[J].中国兽医学报,2017,37(8):1442-1450.
[14]毛玉,刘沅,蔡杰,等.PCV2对小鼠肠道SIgA及相关细胞因子含量的影响[J].中国兽医学报,2017,37(11):2038-2042.
[15] WANG G S,SUN N,TIAN F L,et al.Genetic analysis of porcine circovirus type 2from dead minks[J].J Gen Virol,2016,97(9):2316-2322.
[16] LORINCZ M,CSAGOLA A,BIKSI I,et al.Detection of porcine circovirus in rodents-short communication[J].Acta Vet Hung,2010,58(2):265-268.