码上扫一扫！ |
| 1株谱系7与谱系8.7重组PRRSV的分离鉴定及其基因组特征分析 |
|
陈昊宇1,2, 杨圆1,2, 蓝欣1, 罗程1, 魏维1, 颜一雪1, 岑明升1, 冯钰1, 加洛1, 谢妃敏1, 李雪静1, 陈新悦1, 刘建奎1,3, 魏春华1,3
|
|
1.龙岩学院 生命科学学院，福建 龙岩 364000;2.福建农林大学 动物科学学院，福建 福州 350002;3.福建省家畜传染病防治与生物技术重点实验室，福建 龙岩 364000
|
|
| 摘要: |
| 【目的】明确从福建省规模化猪场分离的猪繁殖与呼吸综合征病毒（PRRSV）野毒株的分子遗传特 征，为PRRSV的防控提供科学依据。【方法】将PRRSV阳性病料的上清滤液接种于猪肺泡巨噬细胞（PAMs）进行病毒的分离鉴定；采用分段RT-PCR扩增方法对分离株进行全基因组扩增，测序后进行序列拼接获得分离毒株的全基 因组序列；使用DNAStar 7.0软件进行核苷酸与氨基酸序列同源性分析，利用MEGA 6.0软件构建系统进化树，采用重组分析软件RDP 4.1与SimPlot 3.5.1对PRRSV基因组进行重组分析。【结果】成功分离获得1株PRRSV，命名为FJWYC-2025，其基因组全长为15 542 bp（不含polyA尾）。同源性分析显示，该毒株全基因组序列与谱系7 PRRSV代表株（Prime Pac、APRRS 和 PRRSGARD）的核苷酸相似性最高，为98.0%-98.4%，与其他谱系PRRSV-2代表株的相似性为84.3%-91.1%。以全基因组和Nsp2基因构建的遗传进化树显示，FJWYC-2025株与谱系7 PRRSV处于同一进化分支；而基于ORF5构建的遗传进化树则显示，该毒株与谱系8.7的HP-PRRSV处于同一分支。氨基酸序 列分析结果显示，FJWYC-2025株Nsp2蛋白表现出与谱系7毒株一致的36 a（a第810-845位）连续插入，而其GP5蛋白关键氨基酸突变与谱系8.7的HP-PRRSV具有一致性。此外，FJWYC-2025株在ORF1b与ORF2间隔区携带PRRSGARD疫苗株独特的23个核苷酸插入序列。重组分析结果显示，FJWYC-2025株是以谱系7 PRRSV PRRS?GARD疫苗株为主要亲本，谱系8.7 HP-PRRSV GD株为次要亲本的重组病毒，重组断裂点位于ORF3；对断裂点前、 后序列构建系统进化树，结果进一步验证了该重组事件的真实性。【结论】分离的PRRSV毒株FJWYC-2025是谱系7毒株和谱系8.7 HP-PRRSV重组产生的新型变异毒株。 |
| 关键词: 猪繁殖与呼吸综合征病毒 PRRSV谱系7 PRRSV谱系8.7 重组病毒 福建省 |
| DOI：10. 13207/j. jnwafu. 2027. 01. 009 |
| 分类号: |
| 基金项目:福建省科技厅引导项目（2025N0012，2021N0032）；中央引导地方科技发展专项（2021L3028）；福建省自然科学基金项目 （2023J01989） |
|
| Isolation，identification and genomic characterization of a recombinant PRRSV strain from lineages 7 and 8.7 |
|
Chen Haoyu1,2, Yang Yuan1,2, Lan Xin1, Luo Cheng1, Wei Wei1, Yan Yixue1, Cen Mingsheng1, Feng Yu1, Jia Luo1, Xie Feimin1, Li Xuejing1, Chen Xinyue1, Liu Jiankui1,3, Wei Chunhua1,3
|
|
1.College of Life Sciences，Longyan University，Longyan，Fujian 364000，China;2.College of Animal Science，Fujian Agriculture and Forestry University，Fuzhou，Fujian 350002，China;3.Fujian Provincial Key Laboratory for the Prevention and Control of Animal Infectious Diseases and Biotechnology，Longyan，Fujian 364000，China
|
| Abstract: |
| 【Objective】This study aims to clarify the molecular genetic characteristics of the wild-type PRRSV strain isolated from a large-scale swine farm in Fujian Province，thereby providing a scientific basis for the prevention and control of PRRSV.【Method】The supernatant filtrate of PRRSV-positive clinical samples was inoculated into porcine alveolar macrophages（PAMs） for virus isolation and identification. The complete genome of the isolate was then amplified using segmented RT-PCR，and the amplified products were cloned and sequenced to obtain the full-length genomic sequence. Homology analysis of nucleotide and amino acid se?quences was performed with DNAStar 7.0 software，while phylogenetic trees were constructed by MEGA 6.0 software. Recombination analysis was carried out employing RDP 4.1 and SimPlot 3.5.1 software.【Result】A PRRSV strain，designated FJWYC-2025，was successfully isolated from PAMs，with a full-length genome of 15 542 bp（excluding the polyA tail）.Homology analysis showed that FJWYC-2025 exhibited the highest nucleotide identity （98.0%-98.4%） with lineage 7 PRRSVs strains （Prime Pac，APRRS，and PRRS?GARD），while sharing 84.3%-91.1% identity with strains of other PRRSV-2 lineages. Phylogenetic analysis based on the full-length genome and Nsp2 gene revealed that FJWYC-2025 clustered with lineage7 PRRSV strains. In contrast，in the ORF5-based tree，it grouped with HP-PRRSV strains of lineage 8.7.Amino acid se?quence analysis revealed that the Nsp2 protein of FJWYC-2025 exhibited a continuous 36-amino-acid insertion （positions 810-845） identical to that of lineage 7 PRRSV strains，whereas the key amino acid mutations in GP5 were consistent with those of lineage 8.7 HP-PRRSV. Additionally，a 23-nucleotide insertion unique to the PRRSGARD vaccine strain was identified in the intergenic region between ORF1b and ORF2 of FJWYC-2025.Recombination analysis showed that FJWYC-2025 was a recombinant virus，with the lineage 7 PRRSV strain PRRSGARD as the major parental and the lineage 8.7 HP-PRRSV strain GD as the minor parental，and one recombination breakpoint was identified within the ORF3. This recombination event was further verified by phylogenetic analysis of segmented genome sequences flanking the breakpoint.【Conclusion】The isolated PRRSV strain FJWYC-2025 is a novel recombinant variant resulting from lineage 7 PRRSV and lineage 8.7 HP-PRRSV. |
| Key words: porcine reproductive and respiratory syndrome virus PRRSV lineage 7 PRRSV lineage 8.7 recombinant virus Fujian Province |
