2021年~2022年山东部分地区猪繁殖与呼吸综合征病毒的遗传进化分析  

Genetic evolution analysis of PRRSV in some areas of Shandong Province from 2021 to 2022

作　　者：吕硕林 于海丽 王彬[1] 时建皓 单虎[1] 马清霞 张传美[1] 杨海燕[1] LV Shuo-lin;YU Hai-li;WANG Bin;SHI Jian-hao;SHAN Hu;MA Qing-xia;ZHANG Chuan-mei;YANG Hai-yan(College of Veterinary Medicine,Qingdao Agricultural University,Qingdao 266109,China)

机构地区：[1]青岛农业大学动物医学院,山东青岛266109

出　　处：《中国预防兽医学报》2023年第12期1207-1215,1222,共10页Chinese Journal of Preventive Veterinary Medicine

基　　金：跨区域科技合作(对口支援)项目(23-1-3-11-zyyd-nsh)。

摘　　要：为了解山东部分地区PRRSV的分子流行及遗传进化情况,本研究对2021年~2022年采自山东部分地区的214份疑似PRRSV感染猪的临床样品通过RT-PCR检测,并经PCR扩增PRRSV阳性样品的ORF5基因和Nsp2基因,采用MegAlign和MEGA X软件对这两个基因进行同源性、遗传进化及其编码的关键氨基酸位点突变及分析。结果显示,214份临床样品中检出51份PRRSV阳性样品,检出率为23.83%。表明山东部分地区PRRSV的感染较为严重。经PCR扩增并测序得到26条ORF5基因序列和20条Nsp2基因序列。26条ORF5基因序列之间的同源性为98.7%~77.8%,与6株PRRSV参考株(经典株VR2332、Ch-1a、疫苗株JXA1、QYYZ及流行株NADC30、NADC34)相应基因序列的同源性在80.1%~98.7%,其中210412等17株PRRSV ORF5基因序列与NADC30株ORF5基因序列的同源性为80.1%~91.9%;进化树结果显示,26条ORF5基因分布于4个谱系中,分别为Lineage1 (220406等22株PRRSV ORF5基因)、Lineage3 (220714和211111株)、Sub Lineage8.7 (HP-PRRSV-Like)(210806株)、Sub Lineage5.1(VR2332-Like)(210629株)谱系;20条Nsp2基因主要分布于3个谱系中,其中220110等13株PRRSV Nsp2基因位于Sub Lineage1.8 (NADC30-Like)谱系,210629株位于Lineage5谱系,与VR2332株遗传距离较近;220714等6株PRRSV Nsp2基因位于Lineage8谱系。上述结果表明,山东部分地区流行的主要为PRRSV NADC30株,属于PRRSV-2,且流行的PRRSV的谱系较复杂,主要以Lineage1、Lineage8及Sub Lineage1.8谱系为主。关键氨基酸位点变异分析结果显示,仅Sub Lineage1.8谱系中的211221和220109株GP5氨基酸序列表现为强毒株特征R^(13)及R^(151);210806株与RespPRRS MLV均为A^(137);除210806株在非中和表位(aa180~aa197)处的变化较为活跃外,其余GP5氨基酸序列与参考株GP5氨基酸序列的变异特征基本一致;在T细胞表位(aa117~aa131)中,Sub Lineage1.8谱系中的多数流行株(包括本研究中的211110等12株)均出现L^(120)S、L^(120)F或I^(125)F、I^(125)L变异。210629株To understand the molecular epidemiology and genetic evolution of PRRSV in some areas of Shandong Province,214 clinical samples suspected to be infected with PRRSV collected there in 2021-2022 were detected by RT-PCR,and the ORF5 and Nsp2 genes of PRRSV were amplified in positive samples by PCR.The resulting sequences were analyzed for the homology,genetic evolution and key amino acid site mutation with the MegAlign and MEGA X sofiwares.The results showed that 51 of 214 clinical samples were detected as PRRSV positive samples and the positive detection rate was 23.83%.It shows that there is PRRSV infection in some areas of Shandong.Twenty-six ORF5 gene sequences and 20 Nsp2 gene sequences were obtained by PCR amplification and sequencing.The homology is 98.7%-77.8%among the 26 ORF5 gene sequences and it is 80.1%-98.7%comparing with the corresponding gene sequences of six PRRSV reference strains(classic strain VR2332,Ch-la,vaccine strain JXA1,QYYZ and epidemic strains NADC30 and NADC34)among which 17 PRRSV ORF5 gene sequences like 210412 strain share 80.1%-91.9%of homology with those of NADC30 PRRSV reference strains.The phylogenetic tree analysis showed that 26 ORF5 genes were distributed in four lineages,namely Lineagel(22 PRRSV ORF5 genes such as 220406 strain),Lineage3(220714 and 21111 strain),Sublineare8.7(HP-PRRSV-Like)(210806 strain)and Sublineare5.1(VR 2332-Like)(210629 strain).Twenty Nsp2 genes are mainly distributed in three lineages,among which 13 PRRSV Nsp2 genes,such as 220110 strain,are located in SubLineagel.8(NADC30-Like)pedigree;210629 strain is located in Lineage5 pedigree,which is close to VR2332.Six PRRSV Nsp2 genes,such as 220714 strain,are located in Lineage8 pedigree.The above results show that PRRSV NADC30 strain belongs to PRRSV-2,and the pedigree of PRRSV is complex,mainly including Lineagel,Lineage8 and SubLineagel.8.The results of mutation analysis of key amino acid sites showed that only 211221 and 220109 strains of PRRSV GP5 protein in SubLineage1.8(NADC30-Like)pedigree showed the cha

关 键 词：猪繁殖与呼吸综合征病毒 遗传进化 ORF5基因 NSP2基因 

分 类 号：S852.65[农业科学—基础兽医学]