摘要
为了了解甘肃地区猪群中猪繁殖与呼吸综合征病毒(PRRSV)的流行及遗传变异情况,采用反转录聚合酶链反应(RT-PCR)方法对2020-2023年收集的猪组织样品进行PRRSV ORF5、NSP2和ORF3基因扩增并测序,通过MEGA软件构建遗传进化树进行遗传变异分析。结果显示,共检测了52份猪临床样品,PRRSV核酸阳性率为61.5%(32/52),成功获得了32条ORF5、15条NSP2和14条ORF3基因序列。ORF5基因遗传进化分析表明,2020—2023年甘肃地区猪群中PRRSV流行毒株中存在美洲型1.5谱系(4株)、1.8谱系(24株)、5.1谱系(2株)、8.7谱系(2株);NSP2基因遗传进化分析表明,获得的15条NSP2序列均为1.8谱系;ORF3基因遗传进化分析表明,获得的14条ORF3序列包括1.5谱系(2株)、1.8谱系(11株)、3.5谱系(1株)。基因核苷酸序列与参考毒株的同源性分别为ORF582.3%~99.7%、NSP285.9%~90.0%和ORF389.8%~95.7%;基因编码氨基酸序列与参考毒株的同源性分别为ORF579.6%~99.0%、NSP283.4%~87.7%和ORF385.1%~94.5%。ORF5基因编码的GP5蛋白的信号肽区、高变区、细胞表位区等区域均存在不同程度的变异,其中5株毒株的第33位氨基酸出现了缺失;15株NSP2蛋白氨基酸序列的缺失模式与NADC30株一致。上述结果表明,当前甘肃地区猪群中PRRSV呈现以1.8谱系毒株为优势的美洲型多谱系共循环流行,这对甘肃地区防控PRRSV提供了重要的流行病学数据。
To understand the genetic variation of porcine reproductive and respiratory syndrome virus(PRRSV)circulating in pig herds in Gansu Province from 2020 to 2023,reverse transcription-polymerase chain reaction(RT-PCR)method was employed to amplify ORF5,NSP2 and ORF3 gene of PRRSV from pig tissue samples.Subsequently,three phylogenetic trees based on ORF5,NSP2 and ORF3 gene were constructed using MEGA software,respectively.The results showed that the positive rate for PRRSV gene of 52clinical samples was 61.5%(32/52),and 32 ORF5 sequences,15 NSP2 sequences and 14 ORF3 sequences were successfully obtained.Phylogenetic analysis based on ORF5 gene showed that the circulating PRRSV-2strains were currently classified as:sublineage 1.5(4 strains),sublineage 1.8(24 strains),sublineage 5.1(2 strains),and sublineage 8.7(2 strains)in Gansu.Phylogenetic analysis based on NSP2 gene showed that all of the 15 NSP2 sequences were belonged to sublineage 1.8.Phylogenetic analysis based on ORF3 gene showed that the 14 ORF3 sequences were belonged to sublineage 1.5(2 strains),sublineage 1.8(11 strains)and sublineage 3.5(1 strain).The nucleotide homology with the reference strains were82.3%to 99.7%(ORF5),85.9%to 90.0%(NSP2)and 89.8%to 95.7%(ORF3),respectively.The amino acid sequence homology between the relative strains and the reference strains were 79.6%to 99.0%(ORF5),83.4%to 87.7%(NSP2)and 85.1%to 94.5%(ORF3),respectively.The GP5 protein,encoded by ORF5 gene,exhibited varying degrees of variation,especially in signal peptide region,high variability region,and cell epitope region,and a deletion of the 33rd amino acid was founded in 5 strains.The patterns of amino acid deletion in 15 NSP2 amino acid sequences were identical to NADC30.In conclusion,PRRSV in Gansu was the simultaneous circulation of multiple lineages with the dominant sublineage 1.8,which provided an important data for the prevention and control of PRRSV in Gansu.
作者
焦守德
张婧
李平花
王健
韩庆彦
张雨星
魏达礼
王方洲
白兴文
卢曾军
马雪青
孙普
JIAO Shoude;ZHANG Jing;LI Pinghua;WANG Jian;HAN Qingyan;ZHANG Yuxing;WEI Dali;WANG Fangzhou;BAI Xingwen;LU Zengjun;MA Xueqing;SUN Pu(State Key Laboratory for Animal Disease Control and Prevention/College of Veterinary Medicine of Lanzhou University/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730000,China;Gansu Province Research Center for Basic Disciplines of Pathogen Biology,Lanzhou 730046,China;Gansu Center for Animal Disease Control and Prevention,Lanzhou 730046,China;Gansu Agricultural University,Lanzhou 730070,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2024年第11期1482-1490,共9页
Chinese Veterinary Science
基金
中国农业科学院创新工程项目(CAAS-ASTIP-2022-LVRI)
国家生猪技术创新中心先导科技项目(NCTIP-XD/C03)
国家生猪技术创新中心牧原项目(NCTIP-MY23004)
甘肃省生猪健康养殖疫病防控及生物安全技术集成试验示范项目(20201007-2)。
