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目的 分析2025年北京市人副流感病毒4型（Human parainfluenza virus type 4,HPIV4）的基因特征。方法 采集2025年北京市流感样病例（Influenza-like illness, ILI）咽拭子标本21002份、严重急性呼吸道感染病例（Severe acute respiratory infection,SARI）咽拭子或下呼吸道标本7570份，进行HPIV4病原体筛查。对检测为HPIV4阳性且Ct值<30的样本进行全基因组测序。采用多重PCR方法扩增HPIV4的全基因组序列，采用BioEdit软件对病毒血凝素-神经氨酸酶（Hemagglutinin-neuraminidase protein,HN）和融合蛋白（Fusion protein,F）基因的核苷酸及氨基酸变异分析，并构建全基因组、HN和F基因的系统发育树，采用NetNGlyc 1.0 Server在线工具预测HPIV4的HN和F蛋白的糖基化位点，并采用SimPlot 3.5.1软件进行重组分析。结果 HPIV4总阳性率为0.31%(81/25872),ILI与SARI病例阳性率分别为0.30%(63/21002)和0.24%(18/7570)。病毒流行高峰主要集中在夏、秋季。测序获得19条HPIV4全基因组序列，其中优势血清亚型为HPIV4b亚型C分支，占比78.95%(15/19),HPIV4a主要为C3分支，占比21.05%(4/19)。19条完整基因组序列（平均测序深度>1100x）长度约为17,300 bp左右，Q30均>85%,GC含量在35.9%～36.3%。HPIV4b的HN、F基因核苷酸同源性分别为98.28%～100.00%、97.81%～100.00%,HPIV4a的HN、F基因核苷酸同源性分别为89.26%～99.93%、99.51%～99.82%。变异分析显示，HPIV4b与HPIV4a在F蛋白发现型别特异性氨基酸变异位点。HN与F基因均受到纯化选择作用。HN蛋白存在5个潜在N-糖基化位点，F蛋白存在3个潜在N-糖基化位点；HN与F基因均未检测到重组事件。结论 2025年北京市HPIV4流行特征呈型别特异性时间分布，HPIV4b为本地优势流行血清亚型；HPIV4基因组存在型别特异性氨基酸变异位点，整体进化以纯化选择为主，未检测到基因重组现象，糖基化位点存在保守性及型别特异性差异。本研究结果为北京市HPIV4的长期监测与防控策略制定提供了重要的基因组数据支撑。
Abstract：Objective To characterize the genetic features of human parainfluenza virus type 4(HPIV4) circulating in Beijing in 2025. Methods A total of 21,002 throat swab samples from influenza-like illness(ILI) cases and 7,570 throat swab or lower respiratory tract specimens from severe acute respiratory infection(SARI) cases were collected in Beijing in 2025 and screened for HPIV4. Whole-genome sequencing was performed on HPIV4-positive samples with cycle threshold(Ct) values <30. The complete viral genomes were amplified using multiplex PCR. Nucleotide and amino acid variations in the hemagglutinin – neuraminidase(HN) and fusion(F) genes were analyzed using MEGA 6.0. Phylogenetic trees based on whole genomes, HN, and F genes were constructed using MEGA 6.0. Potential N-glycosylation sites in HN and F proteins were predicted using the NetNGlyc 1.0 server, and recombination analysis was conducted using SimPlot 3.5.1. Results The overall positivity rate of HPIV4 was 0.31%(81/25,872), with rates of 0.30%(63/21,002) in ILI cases and 0.24%(18/7,570) in SARI cases. Viral activity peaked in summer and autumn. Nineteen complete HPIV4 genomes were obtained. The predominant subtype was HPIV4b, mainly belonging to clade C(78.95%, 15/19), while HPIV4a sequences were primarily assigned to clade C3(21.05%, 4/19). The genomes(average sequencing depth >1,100×) were approximately 17.3 kb in length, with Q30 values >85% and GC content ranging from 35.9% to 36.3%. For HPIV4b, nucleotide identities of the HN and F genes ranged from 98.28%– 100.00% and 97.81% – 100.00%, respectively, whereas for HPIV4a they ranged from 89.26% – 99.93% and 99.51% – 99.82%. Variation analysis identified subtype-specific amino acid substitutions in the F protein. Both HN and F genes were predominantly under purifying selection. Five potential N-glycosylation sites were identified in the HN protein and three in the F protein. No recombination events were detected in either gene. Conclusion In 2025, HPIV4 circulation in Beijing showed subtype-specific temporal patterns, with HPIV4b as the dominant subtype. The HPIV4 genome exhibits subtype-specific amino acid variation, with evolution primarily driven by purifying selection and no evidence of recombination. Glycosylation sites display both conserved and subtype-specific features. These findings provide important genomic data to support long-term surveillance and control strategies for HPIV4 in Beijing.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.260118
中图分类号:R373
引用信息:
[1]高斌,张维嘉,吴丹,等.2025年北京市人副流感病毒4型全基因组分析[J].病毒学报,2026,42(03):775-782.DOI:10.13242/j.cnki.bingduxuebao.260118.
基金信息:
北京市自然科学基金-海淀原始创新联合基金（No.L232073）,题目：流感病毒表面蛋白糖基化修饰对免疫原性的影响研究~~
2026-05-07
2026-05-07
2026-05-07