2017年福建地区PRRSV分子检测及ORF5基因变异分析
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摘要
【目的】研究2017年福建地区中小规模猪场猪繁殖与呼吸综合征病毒(PRRSV)毒株的分子流行情况。【方法】从福建地区中小规模猪场收集83份疑似蓝耳阳性病料,对编码GP5蛋白的ORF5基因进行了RT-PCR检测和遗传进化分析。【结果】采集的83份疑似蓝耳病料中阳性率为73%,测得的序列之间ORF5的核苷酸相似性达80.3%～100.0%,其中所测的各个分支毒株与其所属分支的代表毒株NADC30、GM2和JXA1的核苷酸相似性分别为92.6%～93.9%、89.8%～93.0%和93.4%～99.3%,检测到新分支中新出现的毒株与JXA1的核苷酸相似性为91.0～93.0%。GP5氨基酸比对结果显示不同亚群GP5信号肽区、诱导表位、主要中和表位及高变区等功能区发生了显著变异。【结论】福建地区出现了PRRSV新分支,使福建PRRSV变异种类更加多样化,建议加强PRRSV流行及变异的监测。
【Objective】To study the molecular prevalence of the porcine peproductive and respiratory syndrome virus(PRRSV) strain in small and medium-sized pig farms of Fujian province in 2017.【Method】We collected 83 suspected blue ear positive disease materials from small and medium-sized pig farms in Fujian province. RT-PCR detection and genetic evolution analysis of ORF5 gene which encoded GP5 protein was performed.【Result】The positive rate of 83 suspected blue ear disease samples was 73%, and the nucleotide similarity among the measured ORF5 sequences was 80.3%-100.0%. The nucleotide similarities between tested strains of different branches and the representative strains of those branches, NADC30, GM2 and JXA1, were92.6%-93.9%, 89.8%-93.0% and 93.4%-99.3%, respectively. The new identified strain in the new branch had nucleotide similarity of 91.0%-93.0% to JXA1. The GP5 amino acid alignment showed significant variation in functional regions such as GP5 signal peptide region, induction epitope, principal neutralizing epitope and hypervariable region in different subgroups.【Conclusion】A new branch of PRRSV has emerged in Fujian area, making the PRRSV variants in Fujian more diverse. It is necessary to strengthen the monitoring of PRRSV epidemics and mutations.
【Objective】To study the molecular prevalence of the porcine peproductive and respiratory syndrome virus(PRRSV) strain in small and medium-sized pig farms of Fujian province in 2017.【Method】We collected 83 suspected blue ear positive disease materials from small and medium-sized pig farms in Fujian province. RT-PCR detection and genetic evolution analysis of ORF5 gene which encoded GP5 protein was performed.【Result】The positive rate of 83 suspected blue ear disease samples was 73%, and the nucleotide similarity among the measured ORF5 sequences was 80.3%-100.0%. The nucleotide similarities between tested strains of different branches and the representative strains of those branches, NADC30, GM2 and JXA1, were92.6%-93.9%, 89.8%-93.0% and 93.4%-99.3%, respectively. The new identified strain in the new branch had nucleotide similarity of 91.0%-93.0% to JXA1. The GP5 amino acid alignment showed significant variation in functional regions such as GP5 signal peptide region, induction epitope, principal neutralizing epitope and hypervariable region in different subgroups.【Conclusion】A new branch of PRRSV has emerged in Fujian area, making the PRRSV variants in Fujian more diverse. It is necessary to strengthen the monitoring of PRRSV epidemics and mutations.
引文
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[2]FENG Y,ZHAO T,NGUYEN T,et al.Porcine respiratory and reproductive syndrome virus variants,Vietnam and China,2007[J].Emerg Infect Dis,2008,14(11):1774-1776.
[3]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.
[4]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.
[5]ZHOU Y J,HAO X F,TIAN Z J,et al.Highly virulent porcine reproductive and respiratory syndrome virus emerged in China[J].Transbound Emerg Dis,2008,55(3/4):152-164.
[6]宁宜宝,郑杰,张纯萍,等.我国南方猪高热病的研究:Ⅱ:猪繁殖与呼吸障碍综合征病毒的分离、鉴定和致病性测定[J].中国兽药杂志,2007(1):14-18.
[7]LUNNEY J K,FANG Y,LADINIG A,et al.Porcine reproductive and respiratory syndrome virus(PRRSV):Pathogenesis and interaction with the immune system[J].Annu Rev Anim Biosci,2016,4:129-154.
[8]邓蓉,郑文堂,肖红波.中国生猪生产区域布局与经营规模分析[J].现代化农业,2016(10):44-46.
[9]ZHAO K,YE C,CHANG X,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.
[10]ZHANG Q,JIANG P,SONG Z,et al.Pathogenicity and antigenicity of a novel NADC30-like strain of porcine reproductive and respiratory syndrome virus emerged in China[J].Vet Microbiol,2016,197:93-101.
[11]LIU J K,WEI C H,DAI A L,et al.Complete genomic characterization of two European-genotype porcine reproductive and respiratory syndrome virus isolates in Fujian province of China[J].Arch Virol,2017,162(3):823-833.
[12]BAI X,WANG Y,XU X,et al.Commercial vaccines provide limited protection to NADC30-like PRRSV infection[J].Vaccine,2016,34(46):5540-5545.
[13]张金辉.生猪禁养政策持续加码,养猪企业何去何从?[J].猪业科学,2016(11):142-143.
[14]王亚辉,彭华,李娟.我国21个地区公布畜禽禁养时间表2016年环保政策将更严[J].中国猪业,2016(5):13-18.
[15]刘岳.猪繁殖与呼吸综合征病毒GP5蛋白分段表达及鉴定[D].泰安:山东农业大学,2010.
[16]XIE J,CUI T,CUI J,et al.Epidemiological and evolutionary characteristics of the PRRSV in Southern China from 2010 to 2013[J].Microb Pathog,2014,75:7-15.
[17]SHI M,LAM T T,HON C C,et al.Molecular epidemiology of PRRSV:A phylogenetic perspective[J].Virus Res,2010,154(1/2):7-17.
[18]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(2/3/4):332-340.
[19]申思.HP-PRRSV囊膜糖蛋白5糖基化位点突变及功能初探[D].泰安:山东农业大学,2016.