广东省深圳市屠宰猪群PRRSV ORF5、ORF7、Nsp2基因遗传变异分析OA
Genetic Variation Analysis of PRRSV ORF5,ORF7 and Nsp2 Genes in Pigs for Slaughter in Shenzhen City,Guangdong Province
为明确广东省深圳市屠宰猪群中猪繁殖与呼吸综合征病毒(PRRSV)的隐性带毒状况及流行毒株的遗传变异特征,2023-2024年从深圳市4家生猪屠宰场采集490份淋巴结样品,采用qRT-PCR方法进行PRRSV核酸检测;对阳性样品的ORF5、ORF7和Nsp2基因进行扩增测序,利用MegAlign软件将所得序列与GenBank参考株/流行株进行多序列比对,并构建系统进化树分析遗传进化关系.结果显示:共检出阳性样品69份,其中67份为单一美洲型PRRSV阳性,2份为美洲型与欧洲型PRRSV混合阳性;最终获得19条ORF5、39条ORF7和21条Nsp2基因序列.ORF5的核苷酸和编码氨基酸序列与参考株同源性分别为60.3％～99.67％和51.5％～99.0％,归类为PRRSV-2-Lineage 8/SubLineage 8.7和PRRSV-2-Lineage 1/SubLineage 1.8谱系,对应类JXA1(JXA1-like)和类NADC30(NADC30-like)毒株;ORF7的核苷酸序列与参考株同源性为77.98％～97.98％,大部分也归属于PRRSV-2-Lineage 8/SubLineage 8.7和PRRSV-2-Lineage 1/SubLineage 1.8谱系,为类JXA1 和类NADC30毒株,少部分与PRRSV-2-Lineage 3遗传距离较近;Nsp2的核苷酸序列与参考株同源性为62.97％～99.13％,归类为Lineage 8谱系.此外,由ORF5编码的GP5蛋白多个序列在其信号肽区域发生了氨基酸突变.基于ORF5、ORF7和Nsp2基因的遗传进化分析发现,不同基因对应的毒株谱系分布存在差异.综上所述,深圳市屠宰场屠宰猪群存在PRRSV隐性感染情况,感染毒株以类NADC30为主,且不同毒株间的基因序列存在一定差异,提示可能存在毒株重组事件.本研究为PRRS的精准防控提供了重要的分子流行病学支撑.
In order to clarify the status of subclinical infection with porcine reproductive and respiratory syndrome virus(PRRSV)in pigs for slaughter and the genetic variation characteristics of prevalent strains,490 lymph node samples were collected from 4 slaughterhouses in Shenzhen City from 2023 to 2024,to detect PRRSV nucleic acid using qRT-PCR.The ORF5,ORF7 and Nsp2 genes of positive samples were amplified and sequenced,followed by multiple sequence alignment with reference/epidemic strains registered in GenBank using MegAlign software,and construction of a phylogenetic tree for genetic evolutionary analysis.The results showed that 69 positive samples were detected,including 67 single-positive for American-type PRRSV and 2 co-positive for American-and European-type PRRS V;and a total of 19 ORF5,39 ORF7 and 21 Nsp2 gene sequences were successfully obtained.The nucleotide and encoded amino acid sequences of ORF5 were 60.3％-99.67％and 51.5％-99.0％identical to the reference strain,respectively,and classified into PRRSV-2-Lineage 8/SubLineage 8.7 and PRRSV-2-Lineage 1/SubLineage 1.8 lineages,corresponding to JXA1-like and NADC30-like strains;the nucleotide sequences of ORF7 were 77.98％-97.98％identical to the reference strain,with the majority classified into PRRSV-2-Lineage 8/SubLineage 8.7 and PRRSV-2-Lineage 1/SubLineage 1.8,corresponding to JXAl-like and NADC30-like strains,and the minority closely related to PRRSV-2-Lineage 3.The nucleotide sequence of Nsp2 shared a homology of 62.97％to 99.13％with the reference strain,and was categorized under Lineage 8 lineage.Additionally,amino acid mutations appeared in the signal peptide region of multiple sequences of GP5 protein encoded by ORF5.Phylogenetic analysis based on ORF5,ORF7 and Nsp2 genes revealed discrepancies in lineage distribution of strains corresponding to different genes.In conclusion,PRRSV recessive infection was available in pigs for slaughter in slaughterhouses in Shenzhen City,dominated by NADC30-like strain,and certain genetic differences were available in the genetic sequences of different strains,indicating possible recombination events.Key molecular epidemiological support was provided for targeted prevention and control of PRRS.
陈雨晴;李颖鑫;吴寒光;王润良;陈杰;周建卓;李文萱;柯艳坤
深圳市动植物疫病预防控制中心,广东 深圳 518000深圳市动植物疫病预防控制中心,广东 深圳 518000深圳市动植物疫病预防控制中心,广东 深圳 518000深圳市动植物疫病预防控制中心,广东 深圳 518000深圳市动植物疫病预防控制中心,广东 深圳 518000深圳市动植物疫病预防控制中心,广东 深圳 518000深圳市动植物疫病预防控制中心,广东 深圳 518000深圳市动植物疫病预防控制中心,广东 深圳 518000
农业科技
屠宰场PRRSV遗传进化分析ORF5ORF7Nsp2基因
slaughterhousePRRSVgenetic evolution analysisORF5ORF7Nsp2 gene
《中国动物检疫》 2026 (3)
23-32,94,11
深圳市科技计划资助项目(KCXFZ20230731094003006)
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