谱系1和8重组猪繁殖与呼吸综合征病毒的分离鉴定与遗传进化分析  

Isolation,Identification and Genetic Evolution of a Porcine Reproductive and Respiratory Syndrome Virus Strain Originated from Lineage 1 and 8 Recombination

作　　者：邹舟 黎颖 韦莹莹 刘惠僮 韦祖樟[1,2,3] ZOU Zhou;LI Ying;WEI Ying-ying;LIU Hui-tong;WEI Zu-zhang(Laboratory of Animal infectious Diseases and Molecular Immunology,College of Animal Science and Technology,Guangxi University,Nanning,Guangxi,530005,China;Guangxi Zhuang Autonomous Region Engineering Research Center of Veterinary Biologics,Nanning,Guangxi,530005,China;Guangxi Key Laboratory of Animal Breeding,Disease Control and Prevention,Nanning,Guangxi,530004,China)

机构地区：[1]广西大学动物科学技术学院动物传染病与分子免疫学实验室,广西南宁530005 [2]广西壮族自治区兽用生物制品工程研究中心,广西南宁530005 [3]广西畜禽繁育与疾病防控重点实验室,广西南宁530004

出　　处：《动物医学进展》2025年第1期29-37,共9页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(32172899);国家级大学生创新训练项目(202310593039)。

摘　　要：2023年从广西疑似发生猪繁殖与呼吸综合征病毒(PRRSV)感染猪场的病猪肺脏组织中分离到1株PRRSV,将其命名为GXWZ20230831。采用RT-PCR扩增其全基因组序列并测序,分析遗传进化特点。结果显示,GXWZ20230831能够在Marc-145细胞上增殖,且产生明显的细胞病变;该毒株全基因组长度为15019个核苷酸(nt),与基因2型PRRSV谱系1 NADC30-like毒株的同源性最高,为94.0%,与基因1型PRRSV代表毒株Lelystad virus同源性最低,仅为60.0%;通过Nsp2氨基酸序列比对,结果发现GXWZ20230831存在NADC30-like典型的131(111+1+19)个氨基酸的不连续缺失模式;与美洲型代表株VR2332相比,GXWZ20230831的GP5蛋白在非中和抗原表位发生了V27A、V185A以及R191K 3处突变,同时在GP5蛋白中存在4个潜在的N-糖基化位点;构建的全基因组序列、nsp2基因、ORF5基因遗传进化树均显示该毒株处于谱系1的分支;重组分析表明,GXWZ20230831是1株以谱系1经典毒株NADC30为骨架,谱系8毒株提供重组片段的重组毒株,重组断点位于基因组的nsp1(458-1821 nt)和Nsp4(5405-8179 nt)中。研究结果可为掌握广西地区的PRRSV遗传变异规律提供参考,为制定防控措施提供科学依据。In this study,a porcine reproductive and respiratory syndrome virus(PRRSV)strain was isolated from the lung of a sick pig suspected of PRRSV infection in Guangxi in 2023,and designated as GXWZ20230831.The complete genome was amplified using RT-PCR and sequenced to analyze its genetic evolution.The results showed that GXWZ20230831 could proliferate in Marc-145 cells,exhibiting a marked cytopathic effect(CPE).The length of complete genome was 15019 nucleotides(nt),showing the highest homology(94.0%)with PRRSV-2 NADC30-like strains,while it had the lowest homology(60.0%)with Lelystad virus,the representative strain of PRRSV-1.The alignment of Nsp2 amino acid sequence revealed 131(111+1+19)amino acid discontinuous deletions in GXWZ20230831,characteristic of the NADC30-like deletion pattern.Compared to the representative American strain VR2332,the GP5 protein of GXWZ20230831 exhibited three mutations in non-neutralizing epitopes:V27A,V185A,and R191K.Additionally,there were four potential N-glycosylation sites in the GP5 protein.Phylogenetic analysis of the whole genome sequence,nsp 2 gene,and ORF5 gene indicated that GXWZ20230831 belongs to lineage 1.Recombinant analysis revealed that GXWZ20230831 is a recombinant strain,with NADC30(a classical strain of lineage 1)as the backbone and lineage 8 strain providing the recombinant fragments.The recombination breakpoints were located in nsp1(458-1821 nt)and nsp4(5405-8179 nt)of the genome.These findings provide an important scientific basis for understanding the infection status,genetic variation,and development of prevention and control measures for PRRSV in Guangxi.

关 键 词：猪繁殖与呼吸综合征病毒 分离鉴定 重组 遗传进化分析 

分 类 号：S852.659.6[农业科学—基础兽医学] S858.28[农业科学—兽医学]