1株H1N1亚型猪流感病毒的进化分析及分子特征研究

doi:10.16431/j.cnki.1671-7236.2023.12.030

摘要/Abstract

摘要： 【目的】鉴定贵州省猪流感病毒(Swine influenza virus,SIV)的流行株及其分子生物学特征,为猪流感的病原学研究和流行病学调查提供重要参考。【方法】从贵州省部分养殖场采集猪鼻腔拭子70份,采用RT-PCR和接种犬肾上皮细胞(MDCK)对疑似猪流感的阳性样品进行病毒分离鉴定,对分离株进行克隆测序获得全基因组序列,运用生物信息学在线软件分析毒株的遗传重组情况和关键氨基酸分子特征。【结果】试验分离到1株H1N1亚型SIV,命名为A/swine/Guizhou/ZY/2022(H1N1)。经全基因组测序和遗传进化分析结果显示,该毒株属于G4型欧亚类禽H1N1 SIV。生物信息学在线预测发现,分离株的HA和NA基因属于欧亚类禽H1N1分支,NS基因属于北美重组分支,其余基因片段属于2009年大流行H1N1分支。分离株的HA蛋白裂解位点为PSIQSR↓GL,存在4个潜在糖基化位点,其受体结合位点和抗原位点与欧亚类禽H1N1 SIV高度一致;NA蛋白结构完整,存在5个潜在糖基化位点;分离株PB2蛋白发生了Q⁵⁹¹R、R²⁵¹K突变,PB1蛋白和M2蛋白分别发生了N³⁷⁵S、L⁴⁷³V和S³¹N突变。【结论】从贵州省分离到1株三源重排H1N1亚型SIV,属于G4型欧亚类禽H1N1病毒,其与人源唾液酸α2,6-Gal受体的结合能力可能有所增强,提示需加强对猪流感的流行监测。

关键词: 猪流感病毒; H1N1亚型; 遗传进化; 分子特征

Abstract: 【Objective】 The purpose of this experiment was to identify the circulating strain of Swine influenza virus (SIV) and its molecular biological characteristics in Guizhou province,and provide important reference for the etiological research and epidemiological investigation of swine influenza.【Method】 70 nasal swabs of pigs were collected from some farms in Guizhou province,identified by RT-PCR and inoculated with canine kidney epithelial cells (MDCK)for virus isolation and identification.The whole genome sequence of the strain was obtained by cloning and sequencing.The genetic recombination and key amino acid molecular characteristics of the strain were analyzed by bioinformatics online software.【Result】 The strain of H1N1 subtype SIV was isolated and named A/swine/Guizhou/ZY/2022 (H1N1).Whole genome sequencing and genetic evolution analysis showed that this strain belonged to G4 type Eurasian avian like H1N1 SIV.Bioinformatics online prediction showed that HA and NA genes of the isolated strain belonged to Eurasian avian like H1N1 branch,NS gene belonged to North American recombinant branch,and the rest of the gene fragments belonged to the 2009 pandemic H1N1 branch.The cleavage site of HA protein was PSIQSR↓GL,there were 4 potential glycosylation sites,and the receptor binding sites and antigenic sites were highly consistent with Eurasian avian like H1N1 SIV.The NA protein was structurally intact with 5 potential glycosylation sites.There were Q⁵⁹¹R and R²⁵¹K mutations in PB2 protein,N³⁷⁵S,L⁴⁷³V and S³¹N mutations in PB1 protein and M2 protein,respectively.【Conclusion】 A 3-source rearranged H1N1 subtype SIV strain was isolated from Guizhou province,which belonged to G4 type Eurasian avian like H1N1 virus,and its binding ability to human sialic acid α2,6-GAL receptor might be enhanced,suggesting to strengthen the epidemiological surveillance of swine influenza.

Key words: Swine influenza virus; H1N1 subtype; genetic evolution; molecular characteristics

中图分类号:

S852.65

黄书, 曾智勇, 梁海英, 汤德元, 王彬, 边孟婷, 柳佳佳, 张婧旭, 万娟, 潘向英, 田红利. 1株H1N1亚型猪流感病毒的进化分析及分子特征研究[J]. 中国畜牧兽医, 2023, 50(12): 5084-5093.

HUANG Shu, ZENG Zhiyong, LIANG Haiying, TANG Deyuan, WANG Bin, BIAN Mengting, LIU Jiajia, ZHANG Jingxu, WAN Juan, PAN Xiangying, TIAN Hongli. Evolutionary Analysis and Molecular Characteristics of a H1N1 Subtype Swine Influenza Virus[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(12): 5084-5093.

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