中国兽医杂志2025，Vol.61Issue(3)：58-65,8.DOI:10.20157/j.cnki.zgsyzz.2025.03.008
1株多毒株重组PRRSV的分离鉴定与遗传进化分析
Isolation,Identification and Genetic Evolution Analysis of a Multi-strain Recombinant PRRSV
元娜 1邵明珠 2吕凤霞 3窦丽娜 2李向清 2赵宝凯 2董世山4
作者信息
- 1. 河北农业大学动物医学院,河北 保定 071000||北京大伟嘉生物技术股份有限公司,北京 海淀 100085
- 2. 北京大伟嘉生物技术股份有限公司,北京 海淀 100085
- 3. 河南牧翔动物药业有限公司,河南 郑州 450000
- 4. 河北农业大学动物医学院,河北 保定 071000
- 折叠
摘要
Abstract
To analyze the genetic variation characteristics of porcine reproductive and respiratory syndrome virus(PRRSV),serum samples from sows and lung tissue samples from piglets suspected of being infected with PRRSV at a pig farm in Shanxi Province were tested using real-time fluorescence quantitative reverse transcription PCR(qRT-PCR).Positive lung tissue samples were inoculated into porcine alveolar macrophages(PAMs).After purification by limiting dilution,the PAMs showing cytopathic effects were identified by indirect immunofluorescence assay(IFA).Full-length genome sequencing of the isolated strain was performed by segmental reverse transcription PCR(RT-PCR)amplification,followed by homology and genetic evolution analysis of the open reading frame 5(ORF5)gene and full genome sequence using molecular biological software.Amino acid features of nonstructural protein 2(Nsp2)and recombination analysis of the full genome sequence were also conducted.The results showed that the overall positivity rate of the qRT-PCR was 80%(16/20).One PRRSV strain was isolated from the lung tissue of a PRRSV-positive piglet,named SXZY202301.The genome of this isolated strain was 15 021 bp in length,with a nucleotide homology of 90.4%to the NADC30 strain.It clustered in the same branch of the phylogenetic tree,belonging to Sublineage 1.8.The ORF5 gene of the isolated strain showed a high similarity(95.1%)to the ORF5 gene of the IA/2014/NADC34 strain and clustered in the same branch of the phylogenetic tree,belonging to Sublineage 1.5.The Nsp2 amino acid sequence of the isolated strain exhibited the typical characteristic of 131 amino acid deletions,which is similar to that of the NADC30 strain.Recombination analysis revealed that SXZY202301 had six recombination sites and was a multi-strain recombinant virus,primarily derived from the NADC30-like strain and secondarily from the JXA1 strain and NADC34-like strain.This study provides valuable reference for further research on PRRSV recombination and evolution.关键词
猪繁殖与呼吸综合征病毒(PRRSV)/分离鉴定/测序分析/遗传进化/重组Key words
porcine reproductive and respiratory syndrome virus(PRRSV)/isolation and identification/sequencing analy-sis/genetic evolution/recombination分类
农业科技引用本文复制引用
元娜,邵明珠,吕凤霞,窦丽娜,李向清,赵宝凯,董世山..1株多毒株重组PRRSV的分离鉴定与遗传进化分析[J].中国兽医杂志,2025,61(3):58-65,8.
