高致病性猪繁殖与呼吸综合征病毒疫苗株JXA1-R感染性克隆的构建与鉴定
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摘要
本研究运用反向遗传操作技术平台构建了高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)JXA1-R疫苗株的感染性克隆,经RNA转染BHK-21细胞后成功拯救了病毒,对拯救病毒进行了病毒生物学特性研究,并用该病毒免疫30日龄仔猪进行免疫原性试验。结果表明该感染性克隆与HP-PRRSV JXA1-R疫苗株具有相同的生物学特性,能够很好的诱导猪体产生免疫抗体,并可保护免疫猪抵抗HP-PRRSV NVDC-JXA1强毒株的攻击。为进一步研究HP-PRRSV JXA1-R株疫苗的保护机制以及研发新一代基因工程疫苗奠定了基础。
Based on the reverse genetics technique,the infectious cDNA clone of JXA1-R,an attenuated vaccine strain of the highly pathogenic porcine reproductive and respiratory syndrome virus(HP-PRRSV),was constructed.The chimeric clone was successfully rescued following to be transfected into BHK-21 cells.The rescued viruses maintained the similar biological characteristics as that of the parental virus according to analyse the growth curve of the viruses,30-day-old piglets vaccinated the chimeric viruses can be protected against HP-PRRSV JXA1 strain challenge.All these results are helpful for understanding the protection mechanism of HP-PRRSV JXA1-R strain vaccine and develop a new genetic engineering vaccine.
Based on the reverse genetics technique,the infectious cDNA clone of JXA1-R,an attenuated vaccine strain of the highly pathogenic porcine reproductive and respiratory syndrome virus(HP-PRRSV),was constructed.The chimeric clone was successfully rescued following to be transfected into BHK-21 cells.The rescued viruses maintained the similar biological characteristics as that of the parental virus according to analyse the growth curve of the viruses,30-day-old piglets vaccinated the chimeric viruses can be protected against HP-PRRSV JXA1 strain challenge.All these results are helpful for understanding the protection mechanism of HP-PRRSV JXA1-R strain vaccine and develop a new genetic engineering vaccine.
引文
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[13]翟新验,韩焘,周智.反向遗传学在猪繁殖与呼吸综合征病毒研究中的应用[J].中国兽医杂志,2014(4):52-53.
[14]王晓波,李丽敏,袁万哲,等.猪瘟病毒和猪繁殖与呼吸综合征病毒高致病性/经典毒株多重RT-PCR方法的建立及应用[J].中国兽医学报,2015,35(5):675-679.
[15]LENG X,LI Z,XIA M,et al.Mutations in the genome of the highly pathogenic porcine reproductive and respiratory syndrome virus potentially related to attenuation[J].Vet Microbiol,2012,157(1):50-60.
[16]LI Y,ZHOU L,ZHANG J,et al.Nsp9and Nsp10contribute to the fatal virulence of highly pathogenic porcine reproductive and respiratory syndrome virus emerging in China[J].PLoS Pathogens,2014,10(7):e1004216.
[2]COLLINS J E,BENFIELD D A,CHRISTIANSON W T,et al.Isolation of swine infertility and respiratory syndrome virus(isolate ATCC VR-2332)in North America and experimental reproduction of the disease in gnotobiotic pigs[J].J Vet Diagn Invest,1992,4(2):117-126.
[3]MURTAUGH M P,ELAM M R,KAKACH L T.Comparison of the structural protein coding sequences of the VR-2332and Lelystad virus strains of the PRRS virus[J].Arch Virol,1995,140(8):1451-1460.
[4]ALBINA E.Epidemiology of porcine reproductive and respiratory syndrome(PRRS):an overview[J].Vet Microbiol,1997,55(1):309-316.
[5]TIAN K,YU X,ZHAO T,et al.Emergence of fatal PRRSV variants:unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique hallmark[J].PloS one,2007,2(6):e526.
[6]ZHOU L,CHEN S,ZHANG J,et al.Molecular variation analysis of porcine reproductive and respiratory syndrome virus in China[J].Virus Res,2009,145(1):97-105.
[7]YU X,CHEN N,WANG L,et al.New genomic characteristics of highly pathogenic porcine reproductive and respiratory syndrome viruses do not lead to significant changes in pathogenicity[J].Vet Microbiol,2012,158(3):291-299.
[8]ZHOU L,YANG H.Porcine reproductive and respiratory syndrome in China[J].Virus Res,2010,154(1):31-37.
[9]TIAN Z J,AN T Q,ZHOU Y J,et al.An attenuated live vaccine based on highly pathogenic porcine reproductive and respiratory syndrome virus(HP-PRRSV)protects piglets against HP-PRRS[J].Vet Microbiol,2009,138(1):34-40.
[10]ZHANG S,ZHOU Y,JIANG Y,et al.Generation of an infectious clone of HuN4-F112,an attenuated live vaccine strain of porcine reproductive and respiratory syndrome virus[J].Virol J,2011,8:410.
[11]LENG X,LI Z,XIA M,et al.Evaluation of the efficacy of an attenuated live vaccine against highly pathogenic porcine reproductive and respiratory syndrome virus in young pigs[J].Clin Vaccine Immunol,2012,19(8):1199-1206.
[12]AN T Q,TIAN Z J,ZHOU Y J,et al.Comparative genomic analysis of five pairs of virulent parental/attenuated vaccine strains of PRRSV[J].Vet Microbiol,2011,149(1):104-112.
[13]翟新验,韩焘,周智.反向遗传学在猪繁殖与呼吸综合征病毒研究中的应用[J].中国兽医杂志,2014(4):52-53.
[14]王晓波,李丽敏,袁万哲,等.猪瘟病毒和猪繁殖与呼吸综合征病毒高致病性/经典毒株多重RT-PCR方法的建立及应用[J].中国兽医学报,2015,35(5):675-679.
[15]LENG X,LI Z,XIA M,et al.Mutations in the genome of the highly pathogenic porcine reproductive and respiratory syndrome virus potentially related to attenuation[J].Vet Microbiol,2012,157(1):50-60.
[16]LI Y,ZHOU L,ZHANG J,et al.Nsp9and Nsp10contribute to the fatal virulence of highly pathogenic porcine reproductive and respiratory syndrome virus emerging in China[J].PLoS Pathogens,2014,10(7):e1004216.