河北省PRRSV流行毒株的分离鉴定
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摘要
为了全面调查猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)在河北省的流行情况,本研究共收集河北省833份病死猪临床样品,利用RT-PCR方法对其进行PRRSV检测,共检出419份阳性病料,阳性率高达50.3%。其中NADC30-like PRRSV阳性率为49.1%;HP-PRRSV阳性率为3.1%;经典株PRRSV没有检出;HP-PRRSV和NADC30-like PRRSV共感染样品为16份,共感染率为1.9%。此外,本研究成功地分离到3株NADC30-like毒株,并测通其全基因组序列,与国内外流行毒株进行序列比对和基因重组分析发现,这3个毒株中的QHD1株为重组毒株,亲本可能为PRRSV NADC30毒株和疫苗株RespPRRS MLV,2处重组分别位于7 913～8 015nt和12 780～13 158nt处。本研究为河北省PRRSV的深入研究和防控奠定了基础。
In order to investigate the epidemic situation of porcine reproductive and respiratory syndrome virus(PRRSV)in Hebei Province,833 clinical samples of sick and dead pigs were collected and detected PRRSV by RT-PCR in this study.The results revealed that a total of 419 positive samples were detected,with a positive rate of 50.3%.Thereinto,the positive rate of NADC30-like PRRSV was 49.1%,the positive rate of HP-PRRSV was 3.1%,and the classic strain PRRSV was not detected.The co-infection samples of HP-PRRSV and PRRSV NADC30-like strains were 16 and the co-infection rate was 1.9%.In addition,three NADC30-like strains were successfully isolated and their whole genome were sequenced.Compared with PRRSV epidemic strains through the sequence alignment and gene recombination analysis,the QHD1 isolate was a recombinant strain,and the parent strains may be the PRRSV NADC30 strain and the vaccine strain RespPRRS MLV.The two recombination regions were located at 7 913-8 015 nt and 12 780-13 158 nt,respectively.The research laid the groundwork for the further study on the prevention and control of PRRSV in Hebei Province.
In order to investigate the epidemic situation of porcine reproductive and respiratory syndrome virus(PRRSV)in Hebei Province,833 clinical samples of sick and dead pigs were collected and detected PRRSV by RT-PCR in this study.The results revealed that a total of 419 positive samples were detected,with a positive rate of 50.3%.Thereinto,the positive rate of NADC30-like PRRSV was 49.1%,the positive rate of HP-PRRSV was 3.1%,and the classic strain PRRSV was not detected.The co-infection samples of HP-PRRSV and PRRSV NADC30-like strains were 16 and the co-infection rate was 1.9%.In addition,three NADC30-like strains were successfully isolated and their whole genome were sequenced.Compared with PRRSV epidemic strains through the sequence alignment and gene recombination analysis,the QHD1 isolate was a recombinant strain,and the parent strains may be the PRRSV NADC30 strain and the vaccine strain RespPRRS MLV.The two recombination regions were located at 7 913-8 015 nt and 12 780-13 158 nt,respectively.The research laid the groundwork for the further study on the prevention and control of PRRSV in Hebei Province.
引文
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[14]魏春华,刘建奎,戴爱玲,等.2013-2015年福建省PRRSV ORF3基因亚型遗传变异分析[J].中国兽医学报,2018,38(1):17-24.
[15]WOROBEY M,HOLMES E C.Evolutionary aspects of recombination in RNA viruses[J].J Gen Virol,1999,80(10):2535-2543.
[16]LI Y Y,JI G B,WANG J,et al.Complete genome sequence of an NADC30-Like porcine reproductive and respiratory syndrome virus characterized by recombination with other strains[J].Genome Announc,2016,4(3):e00330-16.
[17]ZHOU L,YANG B N,XU L,et al.Efficacy evaluation of three modified-live virus vaccines against a strain of porcine reproductive and respiratory syndrome virus NADC30-like[J].Vet Microbiol,2017,207:108-116.
[2]CAVANAGH D.Nidovirales:a new order comprising coronaviridae and arteriviridae[J].Arch Virol,1997,142(3):629-633.
[3]TONG G Z,ZHOU Y J,HAO X F,et al.Highly pathogenic porcine reproductive and respiratory syndrome,China[J].Emerg Infect Dis,2007,13(9):1434-1436.
[4]ZHOU L,YANG H C.Porcine reproductive and respiratory syndrome in China[J].Virus Res,2010,154(1/2):31-37.
[5]AN T Q,TIAN Z J,XIAO Y,et al.Origin of highly pathogenic porcine reproductive and respiratory syndrome virus,China[J].Emerg Infect Dis,2010,16(2):365-367.
[6]GUO A J,WU G H,GONG W,et al.Outbreaks of highly pathogenic porcine reproductive and respiratory syndrome in Jiangxi province,China[J].Ir Vet J,2012,65(1):14.
[7]LU W H,TUN H M,SUN B L,et al.Re-emerging of porcine respiratory and reproductive syndrome virus(lineage 3)and increased pathogenicity after genomic recombination with vaccine variant[J].Vet Microbiol,2015,175(2/4):332-340.
[8]ZHOU L,WANG Z C,DING Y P,et al.NADC30-like strain of porcine reproductive and respiratory syndrome virus,China[J].Emerg Infect Dis,2015,21(12):2256-2257.
[9]宋涛.PRRSV遗传变异分析及其nsp2蛋白的定量相互作用组学研究[D].湖北武汉:华中农业大学,2016.
[10]崔丹丹,王傲杰,王新港,等.1株PRRSV类NADC30毒株的分离鉴定及序列分析[J].河南农业科学,2017,46(9):132-138.
[11]魏春华,刘建奎,戴爱玲,等.福建NADC30-like PRRSVFJLY01株的全基因组分子特征分析[J].西北农林科技大学学报(自然科学版),2017,45(3):51-60,67.
[12]ZHAO K,YE C,CHANG X B,et al.Importation and recombination are responsible for the latest emergence of highly pathogenic porcine reproductive and respiratory syndrome virus in China[J].J Virol,2015,89(20):10712-10716.
[13]赵紫印,李春燕,郭抗抗,等.2015年陕西省猪群中4种重要病毒病抗体和病原的检测与分析[J].中国兽医学报,2018,38(5):846-852.
[14]魏春华,刘建奎,戴爱玲,等.2013-2015年福建省PRRSV ORF3基因亚型遗传变异分析[J].中国兽医学报,2018,38(1):17-24.
[15]WOROBEY M,HOLMES E C.Evolutionary aspects of recombination in RNA viruses[J].J Gen Virol,1999,80(10):2535-2543.
[16]LI Y Y,JI G B,WANG J,et al.Complete genome sequence of an NADC30-Like porcine reproductive and respiratory syndrome virus characterized by recombination with other strains[J].Genome Announc,2016,4(3):e00330-16.
[17]ZHOU L,YANG B N,XU L,et al.Efficacy evaluation of three modified-live virus vaccines against a strain of porcine reproductive and respiratory syndrome virus NADC30-like[J].Vet Microbiol,2017,207:108-116.