| 413 | 2 | 165 |
| 下载次数 | 被引频次 | 阅读次数 |
猪繁殖与呼吸综合征病毒（PRRSV）因其高度的遗传变异，导致防控困难，对全球范围内的养猪业造成巨大经济损失。本研究采用猪肺泡巨噬细胞分离获得3株PRRSV类NADC30毒株LY2307、QJ2306和YT2307，其中LY2307可感染Marc-145细胞，QJ2306和YT2307不感染Marc-145细胞。病毒全基因组测序分析结果显示，3株病毒与PRRSV NADC30毒株基因同源性最高，达91.0%～91.6%，全基因组进化树分析显示，3个分离毒株与NADC30毒株位于同一分支，Nsp2蛋白均具有NADC30毒株“111+1+19aa”的缺失模式，GP5蛋白中多个抗原表位存在氨基酸突变。基因重组分析显示，LY2307和QJ2306为NADC30分别与VR-2332和JXA1的重组病毒。生物信息学方法分析病毒Marc-145细胞适应性分子特性，结果显示PRRSV GP2a第78、97和98位氨基酸、GP3第18位氨基酸和GP4第57位氨基酸可能为影响病毒感染Marc-145细胞的关键位点。上述结果表明，本研究3个PRRSV分离毒株均属于类NADC30毒株，但其基因来源、分子特征和细胞适应性存在明显差异，从而加强我国PRRSV流行毒株监测及其分子特征和生物学特性研究，对该病防控和新型疫苗研究具有重要意义。
Abstract：Porcine reproductive and respiratory syndrome virus(PRRSV) exhibits high genetic variability, complicating disease control and causing significant economic losses to the global swine industry. In this study, three PRRSV NADC30-like strains(LY2307, QJ2306, and YT2307) were isolated using porcine alveolar macrophages(PAMs). Among them, LY2307 was able to infect Marc-145 cells, whereas QJ2306 and YT2307 failed to establish infection. Whole-genome sequencing revealed that the three isolates shared the highest genetic homology(91.0%–91.6%) with NADC30 strain. Phylogenetic analysis indicated that all three isolates clustered with NADC30 strains, while the Nsp2 protein exhibited the characteristic "111+1+19 aa" deletion pattern of NADC30 strains. And GP5 protein had multiple amino acid mutations across the antigenic epitopes. Recombination analysis showed that LY2307 and QJ2306 originated from recombination events involving NADC30, VR-2332, and JXA1 strains. Bioinformatic analysis of Marc-145 cell adaptation characteristics identified that the key amino acid residues potentially influencing PRRSV infectivity, are the residues 78, 97, and 98 in GP2a, the residue 18 in GP3, and the residue 57 in GP4. These findings demonstrate that all three PRRSV isolates in this study belong to the NADC30-like strain, yet exhibit distinct genetic origins, molecular characteristics, and cellular adaptability. Enhanced surveillance of PRRSV epidemic strains in China, along with in-depth research on their molecular features and biological properties, holds significant implications for the disease control and the development of novel vaccines in the future.
[1]Wang H, Feng W. Current status of porcine reproductive and respiratory syndrome vaccines[J].Vaccines(Basel), 2024, 12(12):1387. DOI:10. 3390/vaccines12121387.
[2]Kappes MA, Faaberg KS. PRRSV structure,replication and recombination:Origin of phenotype and genotype diversity[J]. Virology, 2015, 479-480:475-486. DOI:10. 1016/j. virol. 2015. 02. 012.
[3]张兴民，张婧，虞凌雪，等.一株多谱系重组NADC34-like-PRRSV的分离鉴定及其基因组特征分析[J].病毒学报，2024, 40(05):1062-1075. DOI:10. 13242/j.cnki. bingduxuebao. 004570.
[4]Yuan N, Yang Z, Lv F, et al. Molecular epidemiology and genetic evolution of porcine reproductive and respiratory syndrome virus in northern China during 2021-2023[J]. Viruses, 2025, 17(1):85. DOI:10. 3390/v17010085.
[5]Zhang J, Bai J, Sun Y, et al. Comparison of pathogenicity of different subgenotype porcine reproductive and respiratory syndrome viruses isolated in China[J]. Microb Pathog, 2022, 168:105607. DOI:10. 1016/j. micpath. 2022. 105607.
[6]冷雪.高致病性猪繁殖与呼吸综合征病毒分离鉴定、遗传变异及致病性分析[D].北京：中国农业科学院，2012.
[7]赵化建.猪繁殖与呼吸综合征病毒遗传变异及JXA1-R疫苗样流行毒株基因特征[D].南京：南京农业大学，2017.
[8]李培勋. 2021-2022年山东省猪繁殖与呼吸综合征流行病学调查及分离毒株遗传进化分析[D].泰安：山东农业大学，2023. DOI:10. 27277/d. cnki.gsdnu. 2023. 000127.
[9]Luo Q, Zheng Y, Zhang H, et al. Research progress on glycoprotein 5 of porcine reproductive and respiratory syndrome virus[J]. Animals(Basel), 2023, 13(5):813. DOI:10. 3390/ani13050813.
[10]Zhou L, Han J, Yang H. The evolution and diversity of porcine reproductive and respiratory syndrome virus in China[J]. Vet Microbiol, 2024, 298:110252. DOI:10. 1016/j. vetmic. 2024. 110252.
[11]李瑞雪，李承辉，陈俭，等.广西南宁猪繁殖与呼吸综合征病毒流行毒株ORF5和NSP2基因的遗传变异分析[J].病毒学报，2025, 41(01):173-183. DOI:10. 13242/j. cnki. bingduxuebao. 004603.
[12]Zhu Z, Yuan L, Hu D, et al. Isolation and genomic characterization of a Chinese NADC34-like PRRSV isolated from Jiangsu province[J]. Transbound Emerg Dis, 2022, 69(4):e1015-e1027. DOI:10. 1111/tbed. 14392.
[13]Li Y, Jiao D, Jing Y, et al. Genetic characterization and pathogenicity of a novel recombinant PRRSV from lineage 1, 8 and 3 in China failed to infect MARC-145cells[J]. Microb Pathog, 2022, 165:105469. DOI:10. 1016/j. micpath. 2022. 105469.
[14]Liu X, Liu X, Bai J, et al. Glyceraldehyde-3-phosphate dehydrogenase restricted in cytoplasmic location by viral GP5 facilitates porcine reproductive and respiratory syndrome virus replication via its glycolytic activity[J].J Virol, 2021, 95(18):e0021021. DOI:10. 1128/JVI. 00210-21.
[15]Liu B, Luo L, Shi Z, et al. Research progress of porcine reproductive and respiratory syndrome virus NSP2 protein[J]. Viruses, 2023, 15(12):2310. DOI:10. 3390/v15122310.
[16]张蕾，洪文姣，张丽琳，等. PRRSV非结构蛋白Nsp2PL2域结构及其去泛素化酶功能[J].病毒学报，2018, 34(4):533-540. DOI:10. 13242/j. cnki.bingduxuebao. 003395.
[17]Zhou L, Zhang J, Zeng J, et al. The 30-amino-acid deletion in the Nsp2 of highly pathogenic porcine reproductive and respiratory syndrome virus emerging in China is not related to its virulence[J]. J Virol, 2009,83(10):5156-5167. DOI:10. 1128/JVI. 02678-08.
[18]Zhang HL, Tang YD, Liu CX, et al. Adaptions of field PRRSVs in Marc-145 cells were determined by variations in the minor envelope proteins GP2a-GP3[J].Vet Microbiol, 2018, 222:46-54. DOI:10. 1016/j.vetmic. 2018. 06. 021.
[19]Chen Y, Huo Z, Jiang Q, et al. The significance of the98th amino acid in GP2a for porcine reproductive and respiratory syndrome virus adaptation in Marc-145 cells[J]. Viruses, 2024, 16(5):711. DOI:10. 3390/v16050711.
[20]Wu X, Qi J, Cong X, et al. Establishment and characterization of a high and stable porcine CD163-expressing MARC-145 cell line[J]. Biomed Res Int,2018, 2018:4315861. DOI:10. 1155/2018/4315861.
[21]Zhu J, He X, Bernard D, et al. Identification of new compounds against PRRSV infection by directly targeting CD163[J]. J Virol, 2023, 97(5):e0005423.DOI:10. 1128/jvi. 00054-23.
基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250095
中图分类号:S852.651
引用信息:
[1]周昊然,苏潼键,张路捷,等.PRRSV类NADC30毒株分离鉴定及Marc-145细胞适应性分子特征分析[J].病毒学报,2025,41(05):1543-1553.DOI:10.13242/j.cnki.bingduxuebao.250095.
基金信息:
国家自然科学基金(项目号：32230103)，题目：猪繁殖与呼吸综合征病毒感染诱导调节性细胞死亡致肺脏损伤的分子机制研究; 国家现代农业产业技术体系专项(项目号：CARS-35); 江苏省农业科技自主创新资金项目(项目号：CX(22)1011)，题目：猪重要传染病新型联合疫苗的创制~~
2025-03-17
2025
2025-05-23
2025-07-23
2025
1
2025-07-24
2025-07-24
2025-07-24