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目的 对云南省纳帕海野鸟栖息地分离获得的1株H9N2亚型禽流感病毒进行全基因组序列分析，阐明其遗传特征、系统进化关系及可能的重配来源，为云南省野鸟中禽流感病毒的分子流行病学研究和跨宿主传播风险评估提供依据。方法 采集野鸟栖息地环境样本，经实时荧光RT-PCR检测和SPF鸡胚分离获得病毒株。采用全基因组RT-PCR扩增与二代测序技术获取8个基因片段序列，利用BLAST进行同源性分析，采用MEGA 7.0软件构建系统进化树，并对HA、NA、NS、M、NP及聚合酶复合体基因的关键氨基酸位点和潜在N-糖基化位点进行分析。结果 2023-2025年共采集野鸟栖息地环境样本647份，检出1株H9N2亚型禽流感病毒，命名为A/Environment/YunnanDiqing/038/2023(H9N2)(DQ038)。全基因组分析显示，DQ038的HA、NA基因属于G9-like谱系，部分内部基因分别归属于F98-like和G1-like谱系，呈现多谱系重配特征。HA蛋白裂解位点为PSRSSRGLF，符合低致病性禽流感病毒特征；受体结合位点存在Q234L（对应H3第226位氨基酸）等突变；未检测到NA抑制剂耐药相关位点突变；PB2基因未出现E627K和D701N突变。结论 纳帕海野鸟栖息地分离的H9N2病毒DQ038具有复杂的多来源、多谱系重配特征，提示野鸟在H9N2病毒跨区域传播与遗传重组中可能发挥重要作用。加强云南高原湿地野鸟禽流感病毒的持续监测，对评估病毒跨宿主传播风险具有重要意义。
Abstract：Objective To characterize the genetic features, phylogenetic relationships, and potential reassortment origins of an H9N2 avian influenza virus isolated from a wild bird habitat in Napahai, Yunnan Province, China, and to provide insights for molecular epidemiological studies and assessment of cross-species transmission risk. Methods Environmental samples were collected from wild bird habitats and screened by realtime reverse transcription PCR(RT-PCR), followed by virus isolation in specific pathogen-free(SPF) embryonated chicken eggs. Whole-genome amplification and next-generation sequencing were performed to obtain sequences of all eight gene segments. Sequence homology was analyzed using BLAST, and phylogenetic trees were constructed using MEGA 7.0. Key amino acid residues and potential N-glycosylation sites in the HA, NA, NS, M, NP, and polymerase complex genes were further analyzed. Results A total of 647 environmental samples were collected from 2023 to 2025, from which one H9N2 avian influenza virus was isolated and designated A/Environment/YunnanDiqing/038/2023(H9N2)(DQ038). Phylogenetic analysis showed that the HA and NA genes of DQ038 belonged to the G9-like lineage, whereas the internal genes clustered within the F98-like and G1-like lineages, indicating a multi-lineage reassortant genotype. The HA cleavage site sequence(PSRSSRGLF) was consistent with low-pathogenic avian influenza viruses. The receptor-binding site harbored the Q234L substitution(corresponding to position 226 in H3 numbering). No amino acid substitutions associated with neuraminidase inhibitor resistance were detected, and no mammalian adaptation – associated mutations(E627K and D701N) were observed in the PB2 gene. Conclusion The H9N2 virus DQ038 isolated from the Napahai wetland exhibits a complex multi-lineage reassortant profile, suggesting that wild birds may play a critical role in the genetic reassortment and regional dissemination of H9N2 viruses. Continuous surveillance of avian influenza viruses in wild bird habitats in Yunnan is essential for evaluating cross-species transmission risk.
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基本信息:
中图分类号:S852.65
引用信息:
[1]刘照生,伏晓庆,罗春蕊,等.云南省纳帕海野鸟栖息地1株H9N2亚型禽流感病毒的全基因组序列分析[J].病毒学报().
基金信息:
云南省疾病预防控制中心科技计划项目（项目号：2023ZX003）,题目：2023-2025年云南省野鸟栖息地外环境禽流感病毒监测;云南省疾病预防控制中心科研项目（项目号：YNAPM2025-003）,题目：呼吸道及肠道传染病感染谱及人群流行特征研究
2026-05-29
2026-05-29
2026-05-29