1株重组类NADC34猪繁殖与呼吸综合征病毒分离鉴定与基因组特征分析  

Isolation and genome analysis of a recombinant NADC34-like porcine reproductive and respiratory syndrome virus

作　　者：刘野 李文忠 杨婧 程宁 杨程 王凯月 吴庆东 张乐 赵瑞利 孙英峰 LIU Ye;LI Wenzhong;YANG Jing;CHENG Ning;YANG Cheng;WANG Kaiyue;WU Qingdong;ZHANG Le;ZHAO Ruili;SUN Yingfeng(College of Animal Science and Veterinary Medicine,Tianjin Agricultural University,Tianjin 300384,China;Tianjin Jinghai Institute of Agricultural Science and Technology,Tianjin 301699,China;Tianjin Jinghai District Agricultural Development Service Center,Tianjin 301699,China)

机构地区：[1]天津农学院动物科学与动物医学学院,天津300384 [2]天津市静海区农业科学技术研究所,天津301699 [3]天津市静海区农业发展服务中心,天津301699

出　　处：《中国兽医学报》2023年第11期2212-2220,共9页Chinese Journal of Veterinary Science

基　　金：天津市科技支撑计划重点资助项目(22YFZCSN00100);天津市优秀企业科技特派员资助项目(22YDTPJC00420);天津市优秀农业科技特派员资助项目(22ZYCGSN00570)。

摘　　要：为分离天津地区猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)并分析其分子遗传进化关系,对来自该地区某猪场疑似PRRSV感染的样品进行RT-PCR检测,阳性样品接种猪肺泡巨噬细胞(PAMs),将出现细胞病变的PAMs进行有限稀释后进行间接免疫荧光鉴定,利用RT-PCR扩增分离纯化株全基因组序列并进行同源性及遗传进化分析,并对分离株进行重组分析和细胞嗜性试验。结果显示,分离获得1株PRRSV毒株(命名为TJbc2021),分离毒株不能感染Marc-145细胞;该毒株全基因组长为15 111 bp,与NADC34的核苷酸同源性最高,为95.2%,属于目前流行于中国的类NADC34 PRRSV,其Nsp2存在100个氨基酸缺失的分子特征;重组分析显示该毒株以谱系1.5毒株(NADC34-like)为主要亲本,以谱系1.8毒株(NADC30-like)为次要亲本,在14 082~14 635 nt(ORF6~ORF7)发生了基因重组。结果表明,分离并鉴定了1株重组类NADC34 PRRSV,可为该地区猪繁殖与呼吸综合征的防控提供参考。To isolate the porcine reproductive and respiratory syndrome virus(PRRSV) in Tianjin and analyze its molecular genetic evolution,RT-PCR tests were performed on samples suspected of PRRSV infection from a pig farm in the area.Positive samples were used to infect porcine alveolar macrophages(PAMs),and PAMs with cytopathic effect(CPE)were identified by indirect immunofluorescence(IFA)after limited dilution.RT-PCR was used to isolate and purify the whole genome sequence of the strain and homology and genetic evolution analysis were performed.Recombinant analysis and cellular tropism tests were performed on the isolate.The results showed that the isolated PRRSV strain(named TJbc2021) did not infect Marc-145 cells.The whole genome length of this strain was 15 111 bp,which has the highest nucleotide homology with NADC34 with a nucleotide homology of 95.2%.Analysis revealed the strain belongs to the NADC34-like PRRSV that is currently mainstream in China,and its Nsp2 has the molecular characteristics of 100 amino acid deletions.Recombinant analysis showed that the strain had genetic recombination at 14 082-14 635nt(ORF6-ORF7)with lineage 1.5strain(NADC34-like)as the main parent and lineage 1.8strain(NADC30-like)as the secondary parent.The results of this study provide reference for the prevention and control of reproductive and respiratory syndrome in pigs.

关 键 词：猪繁殖与呼吸综合征病毒 类NADC34 分离鉴定 重组 细胞嗜性 

分 类 号：S852.65[农业科学—基础兽医学]