2022-2023年无锡市甲型H3N2流感病毒全基因组特征分析
Analysis of the whole genome characteristics of influenza A(H3N2)virus in Wuxi city from 2022 to 2023
徐勇 1王瑞 2於淳安 3鲍静 3周琪 3肖勇 3李泓 1石晓娈 1马广源3
作者信息
- 1. 无锡市滨湖区疾病预防控制中心,无锡 214072
- 2. 南京医科大学,南京 211166
- 3. 无锡市疾病预防控制中心(南京医科大学附属无锡疾病预防控制中心),无锡 214023
- 折叠
摘要
目的 了解无锡市2022-2023年甲型H3N2流感病毒的全基因组和遗传进化特征.方法 采用实时荧光定量RT-PCR方法对流感样病例呼吸道样本进行分型检测,对甲型H3N2流感病毒核酸阳性的样本进行病毒分离培养,红细胞凝集试验(HA)≥1∶8的毒株提取核酸,扩增全基因组序列,构建文库,采用MiSeq测序仪上机测序.以NC_007366.1为参考株,使用CLC Genomics Workbench(Version 23)软件分析下机数据.采用MEGA 7.0软件构建系统进化树,NetNGlyc 1.0 Server软件预测N-糖基化位点.结果 2022-2023年35株甲型H3N2流感病毒流行株之间的核苷酸同源性为96.4％-100％,氨基酸同源性为95.2％～100％.2022年16株流行株属于3C.2a1b.2a.1a进化分支,2023年19株流行株属于3C.2a1b.2a.2a.3a.1进化分支.2022年流行株与对应疫苗株比较HA基因核苷酸序列存在7个差异位点,共享15个突变位点;2023年流行株与对应疫苗株比较HA基因核苷酸序列存在28个差异位点,共享17个突变位点.35株流行株HA基因均缺少N-糖基化位点61:NSS,而2023年19株流行株HA基因新增N-糖基化位点110:NSS.结论 2022年和2023年甲型H3N2流感病毒的HA和NA基因分别属于2个进化分支,与相应疫苗株比较均出现特异性氨基酸位点改变.2022年流行株与疫苗株比较抗原匹配性较好,2023年流行株与疫苗株比较存在抗原匹配性较低的现象.
Abstract
Objective To understand the whole genome and genetic evolution characteristics of the first epidemic influenza A(H3N2)viruses in Wuxi from 2022-2023.Methods Real time fluorescence quantitative RT-PCR method was used to perform typing on respiratory samples of influenza cases.Virus isolation was performed on samples with positive nucleic acid of subtype A H3N2 influenza virus detected.After cell culture,nucleic acid was extracted from strains with red blood cell agglutination test(HA)≥ 1∶8,whole genome sequence was amplified,library was constructed,and computer sequencing was performed using MiSeq sequencer.Using NC_007366.1 as reference strain,the data were analyzed using CLC Genomics Workbench(Version 23)software.The phylogenetic tree was constructed using MEGA 7.0 software,and the N-glycosylation sites were predicted by NetNGlyc 1.0 Server software.Results The nucleotide homology and amino acid homology among 35 strains of influenza A H3N2 virus from 2022 to 2023 were 96.4％-100％and 95.2％-100％,respectively.The 16 epidemic strains in 2022 belong to the 3C.2a1b.2a.1a evolutionary branch,while the 19 epidemic strains in 2023 belong to the 3C.2a1b.2a.2a.3a.1 evolutionary branch.There are 7 differences in the nucleotide sequence of the HA gene between the 2022 epidemic strain and the corresponding vaccine strain,sharing 15 mutation sites;There are 28 differences in the nucleotide sequence of the HA gene between the 2023 epidemic strain and the corresponding vaccine strain,sharing 17 mutation sites.The HA genes of 35 epidemic strains all lack N-glycosylation site 61:NSS,while in 2023,the HA genes of 19 epidemic strains added N-glycosylation site 110:NSS.Conclusions The HA and NA genes of influenza A H3N2 virus in 2022 and 2023 belong to two evolutionary branches,respectively,and both show specific amino acid site changes compared to the corresponding vaccine strains.The antigen matching between the 2022 epidemic strain and the vaccine strain is relatively good,while there is a risk of low antigen matching between the 2023 epidemic strain and the vaccine strain.
关键词
甲型H3N2流感病毒/全基因组测序/变异/N-糖基化Key words
Influenza A H3N2 virus/Whole genome sequencing/Variation/N-Glycosylation引用本文复制引用
基金项目
无锡市医学发展学科项目(FZXK2021010)
出版年
2024
NETL
NSTL
万方数据