2020—2024年广西H3N2亚型禽流感病毒HA和NA基因分子特征分析

doi:10.16431/j.cnki.1671-7236.2025.09.032

中国畜牧兽医 ›› 2025, Vol. 52 ›› Issue (9): 4358-4367.doi: 10.16431/j.cnki.1671-7236.2025.09.032

• 预防兽医 • 上一篇

2020—2024年广西H3N2亚型禽流感病毒HA和NA基因分子特征分析

尹彦文¹, 熊陈勇², 韦伟¹, 施开创¹, 郑敏¹, 崔鹏飞³, 韦显凯¹, 李致远⁴, 刘惠心⁴, 冯淑萍¹, 卢春花¹, 屈素洁¹, 陆文俊¹, 李雪珍⁵, 王睿敏⁶, 邓国华³, 欧长波⁴

1. 广西动物疫病预防控制中心, 南宁 530001;
2. 广西农垦永新畜牧集团龙北牧业有限公司, 崇左 532400;
3. 中国农业科学院哈尔滨兽医研究所, 兽医生物技术国家重点实验室, 农业农村部动物流感重点实验室, 哈尔滨 150069;
4. 广西大学动物科学技术学院, 南宁 530005;
5. 百色市动物疫病预防控制中心, 百色 533000;
6. 广西水牛研究所, 南宁 530001

收稿日期:2024-12-14 发布日期:2025-08-29
通讯作者: 邓国华, 欧长波 E-mail:dgh1971@163.com;ouchangbo@gxu.edu.cn
作者简介:尹彦文，E-mail：yanwen0349@126.com；熊陈勇，E-mail：1369400342@qq.com。
基金资助:
广西科技重大专项(桂科AA23062050)；国家现代农业产业体系广西家禽产业创新团队(nycytxgxcxtd-2024-19)

Molecular Characterization of HA and NA Genes of H3N2 Subtype Avian Influenza Viruses in Guangxi During 2020 to 2024

YIN Yanwen¹, XIONG Chenyong², WEI Wei¹, SHI Kaichuang¹, ZHENG Min¹, CUI Pengfei³, WEI Xiankai¹, LI Zhiyuan⁴, LIU Huixin⁴, FENG Shuping¹, LU Chunhua¹, QU Sujie¹, LU Wenjun¹, LI Xuezhen⁵, WANG Ruimin⁶, DENG Guohua³, OU Changbo⁴

1. Guangxi Center for Animal Disease Control and Prevention, Nanning 530001, China;
2. Guangxi State Farms Yongxin Animal Husbandry Group Longbei Animal Husbandry Co., Ltd., Chongzuo 532400, China;
3. Animal Influenza Laboratory of Ministry of Agriculture and Rural Affairs, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China;
4. College of Animal Science and Technology, Guangxi University, Nanning 530005, China;
5. Baise Center for Animal Disease Control and Prevention, Baise 533000, China;
6. Guangxi Buffalo Research Institute, Nanning 530001, China

Received:2024-12-14 Published:2025-08-29

摘要/Abstract

摘要： 【目的】探究H3N2亚型禽流感病毒(AIV)广西毒株分子遗传进化特征。【方法】采集2020—2024年广西各地的家禽咽拭子和肛拭子病料，采用实时荧光定量RT-PCR方法鉴别检测H3N2亚型AIV，选取部分阳性样本经尿囊腔接种SPF鸡胚，收集鸡胚尿囊液，提取核酸后通过PCR扩增血凝素(HA)和神经氨酸酶(NA)基因并测序，进行序列相似性、系统发育、氨基酸变异位点及遗传进化速率分析。【结果】共获得43株HA和NA基因序列，编码区大小为1 701和1 410 bp。相似性比对分析显示，本研究获得的HA和NA基因之间核苷酸序列相似性分别为86.0%～99.9%和89.7%～99.9%，与GenBank中禽源、人源、猪源和犬源核苷酸序列相似性分别为78.4%～99.7%和78.1%～99.1%。广西H3N2亚型AIV毒株HA和NA基因属于欧亚谱系，与禽源亲缘关系最近，与人源/猪源最远。HA蛋白在第18—20、22—24、38—40、54—56、61—63、181—183、301—303和499—501位氨基酸处出现糖基化，NA蛋白在第61—63、69—71、86—88、143—145、146—148、200—202、234—236、313—315和402—404位氨基酸处出现糖基化，部分氨基酸位点发生变异；HA蛋白裂解位点为340PEKQTR↓GLF348和340PERQTR↓GLF348。遗传进化速率估算结果显示，HA和NA基因遗传进化速率分别为3.26×10^-3和3.67×10^-3 替换/(位点·年)，HA基因种群规模在不同时间段表现出不同的动态进化特征。【结论】 H3N2亚型AIV广西流行毒株与同种流行毒株亲缘性较近，部分HA和NA蛋白发生特有氨基酸突变，不同种属毒株进化趋势不一致，具有遗传多样性特征。试验结果为防控H3N2亚型AIV提供基础数据。

关键词: 禽流感病毒; H3N2亚型; 遗传进化; 相似性

Abstract: 【Objective】 The experiment was conducted to explore the molecular genetic evolution characteristics of H3N2 subtype Avian influenza virus (AIV) strains in Guangxi. 【Method】 Disease samples of poultry throat swabs and anal swabs from various places in Guangxi from 2020 to 2024 were collected,and the H3N2 subtype AIV was identified and detected by Real-time quantitative RT-PCR.Some positive samples were selected and inoculated into SPF chicken embryos through the allantoic cavity,and the allantoic fluid of chicken embryos was collected.After extracting nucleic acids,hemagglutinin (HA) and neuraminidase (NA) genes were amplified by PCR and sequenced,and analyses of sequence similarity,phylogenetic,amino acid variation sites and genetic evolution rate were conducted. 【Result】 A total of 43 HA and NA gene sequences were obtained,with the coding region sizes of 1 701 and 1 410 bp,respectively.The similarity comparison analysis showed that the nucleotide sequence similarities between HA and NA genes obtained in this study were 86.0%-99.9% and 89.7%-99.9%,respectively.The similarities of nucleotide sequences with avian,human,porcine and canine sources in GenBank were 78.4%-99.7% and 78.1%-99.1%,respectively.HA and NA genes of H3N2 subtype AIV in Guangxi belonged to the Eurasian lineage,which was most closely related to avian origin and most closely related to human/porcine origin.Glycosylation occured at amino acids 18-20,22-24,38-40,54-56,61-63,181-183,301-303 and 499-501 of HA protein，and occured at amino acids 61-63,69-71,86-88,143-145,146-148,200-202,234-236,313-315 and 402-404 of NA protein.Some amino acid sites had undergone variations.HA protein was cleaved at sites 340PEKQTR↓GLF348 and 340PERQTR↓GLF348.Genetic evolution rate estimation showed that the genetic evolution rates of HA and NA genes were 3.26×10^-3 and 3.67×10^-3 substitutions/(site·year),respectively,and the population size of HA gene showed different dynamic evolution characteristics in various periods. 【Conclusion】 The current epidemic strains of H3N2 subtype AIV in Guangxi were closely related to the homologous epidemic strains,some HA and NA proteins havd unique amino acid mutations,and the evolutionary trend of strains of different genera was inconsistent with the characteristics of genetic diversity,which providing basic data for prevention and control of H3N2 subtype AIV.

Key words: Avian influenza virus; H3N2 subtype; genetic evolution; similarity

中图分类号:

S852.65^＋9.2

尹彦文, 熊陈勇, 韦伟, 施开创, 郑敏, 崔鹏飞, 韦显凯, 李致远, 刘惠心, 冯淑萍, 卢春花, 屈素洁, 陆文俊, 李雪珍, 王睿敏, 邓国华, 欧长波. 2020—2024年广西H3N2亚型禽流感病毒HA和NA基因分子特征分析[J]. 中国畜牧兽医, 2025, 52(9): 4358-4367.

YIN Yanwen, XIONG Chenyong, WEI Wei, SHI Kaichuang, ZHENG Min, CUI Pengfei, WEI Xiankai, LI Zhiyuan, LIU Huixin, FENG Shuping, LU Chunhua, QU Sujie, LU Wenjun, LI Xuezhen, WANG Ruimin, DENG Guohua, OU Changbo. Molecular Characterization of HA and NA Genes of H3N2 Subtype Avian Influenza Viruses in Guangxi During 2020 to 2024[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(9): 4358-4367.

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2020—2024年广西H3N2亚型禽流感病毒HA和NA基因分子特征分析

Molecular Characterization of HA and NA Genes of H3N2 Subtype Avian Influenza Viruses in Guangxi During 2020 to 2024

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