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Article(id=1324078120709534234, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1324078037817504735, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202508170, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1755014400000, receivedDateStr=2025-08-13, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1793614424159, onlineDateStr=2026-11-02, pubDate=1770652800000, pubDateStr=2026-02-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793614424159, onlineIssueDateStr=2026-11-02, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793614424159, creator=13701087609, updateTime=1793614424159, updator=13701087609, issue=Issue{id=1324078037817504735, tenantId=1146029695717560320, journalId=1227665162245664772, year='2026', volume='53', issue='3', pageStart='385', pageEnd='576', issueExtLink='null', onlineDate='null', pubDate='1770652800000', pubDateStr='2026-02-10', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793614404396, creator='13701087609', updateTime=1793614474144, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1324078330420527919, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1324078037817504735, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1324078330420527920, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1324078037817504735, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=533, endPage=538, ext={EN=ArticleExt(id=1324078120940220955, articleId=1324078120709534234, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Molecular surveillance of H3N8 avian influenza virus in live poultry markets associated with human cases in Changsha City, 2022—2023, columnId=1228016573156360233, journalTitle=Modern Preventive Medicine, columnName=Disease Control and Prevention, runingTitle=null, highlight=null, articleAbstract=
Objective

To characterize H3N8 virus contamination and molecular evolution in poultry and environmental samples from the live poultry markets associated with Hunan Province’s first human H3N8 avian influenza virus (AIV) case, providing evidence for avian influenza epidemic prevention and control.

Methods

Active surveillance for H3N8 avian influenza virus was implemented over 17 months (April 2022—December 2023) in live poultry markets associated with Hunan Province’s first human case. We collected 837 samples, including poultry oropharyngeal/cloacal swabs, feces, air, sewage, drinking water, cage swabs, and other surface swabs. Samples were screened for H3N8 nucleic acid using real-time PCR. Positive specimens underwent high-throughput sequencing. Hemagglutinin (HA) and neuraminidase (NA) gene sequences were analyzed by BLAST, with amino acid alignment and phylogenetic tree construction performed using BioEdit and MEGA-X, respectively.The statistical analysis method for nucleic acid test results was Fisher’s exact test.

Results

The overall H3N8 nucleic acid positivity rate was 2.63% (22/837). The highest positivity rate (18.37%) occurred in the month of the human case (May 2022),significantly higher than in the other 16 months (χ2=49.159, P<0.001). Viral nucleic acid was detected in six sample types, with cage swabs showing the highest positivity rate (8.43%, χ2=14.437, P=0.011). Three H3N8 viruses were successfully sequenced from poultry drinking water and chicken oropharyngeal/cloacal swabs. Their HA and NA genes exhibited the highest nucleotide similarity (99.71-99.89%) to the human-origin strain (A/China/CSKFQ-22-5/2022). The HA protein cleavage site (residues 340-348) was PEKQTRGLF, and the receptor-binding site residues (242 and 244) were Q and G, indicating low pathogenicity and avian receptor-binding specificity. No neuraminidase inhibitor resistance mutations (H273Y, R291K) were detected in the NA protein. Phylogenetic analysis revealed that the HA and NA genes of all three viruses belonged to the Eurasian lineage and formed a distinct subclade with H3N8 viruses of human, chicken, and environmental origin from China.

Conclusion

The level of H3N8 virus contamination in live poultry markets shows an association with human infection cases. Enhanced surveillance of H3N8 virus contamination in these markets is warranted.

, authors=Ru-sheng ZHANG1, Hui-yuan FU1, Rui-qing PEI1, Zheng HUANG1, Can LUO1, Jing-fang CHEN1, Shan CHEN1, La-mei CHEN2, authorsList=Ru-sheng ZHANG, Hui-yuan FU, Rui-qing PEI, Zheng HUANG, Can LUO, Jing-fang CHEN, Shan CHEN, La-mei CHEN, authorCompany=null, correspAuthors=La-mei CHEN, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1324078123079315998, articleId=1324078120709534234, tenantId=1146029695717560320, journalId=1227665162245664772, language=CN, title=2022—2023年长沙市病例涉疫活禽市场中H3N8禽流感病毒分子监测, columnId=1228016573844226129, journalTitle=现代预防医学, columnName=疾病预防控制, runingTitle=null, highlight=null, articleAbstract=
目的

分析湖南省首例人感染H3N8亚型禽流感病毒（avian influenza virus, AIV）（简称H3N8病毒）病例涉疫活禽市场中H3N8病毒污染状况及其分子进化特征，为禽流感疫情防控提供依据。

方法

于2022年4月—2023年12月（共17个月）对湖南省首例人感染H3N8病例的涉疫活禽市场进行H3N8病毒监测，采集家禽口咽/泄殖腔拭子、粪便、空气、污水、饮水、笼具涂抹物及其他物体表面涂抹样本共837份。采用Real-Time PCR技术检测样本中H3N8病毒核酸，对阳性样本进行高通量核苷酸测序，对获得的病毒血凝素（hemagglutinin, HA）和神经氨酸酶（neuraminidase, NA）基因序列进行BLAST分析，利用BioEdit和Mega X软件分别进行氨基酸序列比对及系统进化树构建。核酸检测结果统计学分析方法采用Fisher精确检验。

结果

样本总体H3N8病毒核酸阳性率为2.63% (22/837)。病例发生当月（2022年5月）阳性率最高，为18.37%，显著高于其他16个月份（χ2=49.159, P<0.001）。6类样本中检出病毒核酸，其中笼具涂抹样本阳性率最高，为8.43%（χ2=14.437, P=0.011）。测序获得3份来源于家禽饮水和鸡口咽/泄殖腔拭子样本的H3N8病毒，其HA、NA基因与人源株（A/China/CSKFQ-22-5/2022）核苷酸相似性最高（99.71～99.89%）。HA蛋白裂解位点（第340～348位氨基酸）为PEKQTRGLF，受体结合位点（第242、244位氨基酸）为Q、G，呈现低致病性及禽类受体结合特征。NA蛋白未发现神经氨酸酶抑制剂耐药位点（H273Y、R291K）突变。系统进化分析显示，3株病毒的HA、NA基因均属欧亚分支，并与国内人、鸡及环境来源的H3N8病毒共同形成一个独立进化分支。

结论

活禽市场H3N8病毒污染程度与人感染病例的发生存在一定相关性，应密切监测活禽市场中的H3N8病毒污染状况。

, authors=张如胜1, 扶会媛1, 裴瑞青1, 黄政1, 罗璨1, 陈静芳1, 陈姗1, 陈腊梅2, authorsList=张如胜, 扶会媛, 裴瑞青, 黄政, 罗璨, 陈静芳, 陈姗, 陈腊梅, authorCompany=null, correspAuthors=陈腊梅, authorNote=

张如胜（1977-），男，硕士，主任技师，研究方向：新发传染病的分子流行病学研究

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陈腊梅，E-mail:
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张如胜（1977-），男，硕士，主任技师，研究方向：新发传染病的分子流行病学研究

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张如胜（1977-），男，硕士，主任技师，研究方向：新发传染病的分子流行病学研究

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Journal of Medical Virology, 2023, 95(7): e28912., articleTitle=Biological features of human influenza A(H3N8)viruses in China, refAbstract=null)], funds=[Fund(id=1324078128972313185, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, awardId=2023JJ60399; 2023JJ60070, language=CN, fundingSource=湖南省自然科学基金(2023JJ60399; 2023JJ60070), fundOrder=null, country=null), Fund(id=1324078129060393570, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, awardId=R2023169, language=CN, fundingSource=湖南省卫生健康高层次人才重大科研专项(R2023169), fundOrder=null, country=null), Fund(id=1324078129127502435, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, awardId=202203094936, language=CN, fundingSource=湖南省卫生健康委科研项目(202203094936), fundOrder=null, country=null), Fund(id=1324078129211388516, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, awardId=KJ-B2023099, language=CN, fundingSource=长沙市卫健委科研项目(KJ-B2023099), fundOrder=null, country=null), Fund(id=1324078129299468901, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, awardId=kq2403183, language=CN, fundingSource=长沙市自然科学基金项目(kq2403183), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1324078123280642591, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, xref=1., ext=[AuthorCompanyExt(id=1324078123289031200, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, companyId=1324078123280642591, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Changsha Center for Disease Control and Prevention (Changsha Municipal Health Comprehensive Supervision and Law Enforcement Bureau), Changsha, Hunan 410007, China), AuthorCompanyExt(id=1324078123297419809, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, companyId=1324078123280642591, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.长沙市疾病预防控制中心(长沙市卫生综合监督执法局)，湖南 长沙 410007)]), AuthorCompany(id=1324078123364528674, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, xref=2., ext=[AuthorCompanyExt(id=1324078123372917283, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, companyId=1324078123364528674, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.开福区疾病预防控制中心(开福区卫生综合监督执法局)，湖南 长沙 410007)])], figs=[ArticleFig(id=1324078128011817559, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, language=EN, label=Figure 1, caption=Phylogenetic analysis of the HA and NA genes of the H3N8 viruses isolates from the epidemic-related live poultry markets, figureFileSmall=Y2nzCNxRCwW+tLy8vQip4Q==, figureFileBig=uRhfxaJokrLG1bfDFWMeaQ==, tableContent=null), ArticleFig(id=1324078128087315032, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, language=CN, label=图1, caption=病例涉疫活禽市场H3N8病毒HA和NA基因进化分析

注：本研究的H3N8病毒标记为▲；人源H3N8病毒标记为●；A为HA基因；B为NA基因。

, figureFileSmall=Y2nzCNxRCwW+tLy8vQip4Q==, figureFileBig=uRhfxaJokrLG1bfDFWMeaQ==, tableContent=null), ArticleFig(id=1324078128305418841, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, language=EN, label=Table 1, caption=

H3N8 virus nucleic acid detection results from the epidemic-related live poultry markets at different monitoring time points

, figureFileSmall=null, figureFileBig=null, tableContent=
监测月份（年/月）检测数（份）H3N8阳性标本数(阳性率，%)
2022/04431（2.33）
2022/059818（18.37）
2022/06461（2.17）
2022/08470（0）
2022/09471（2.13）
2022/11471（2.13）
2023/02460（0）
2023/03470（0）
2023/04450（0）
2023/05470（0）
2023/06460（0）
2023/07450（0）
2023/08460（0）
2023/09460（0）
2023/10470（0）
2023/11460（0）
2023/12480（0）
合计83722(2.63)
), ArticleFig(id=1324078128385110618, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, language=CN, label=表1, caption=

病例涉疫活禽市场不同监测时间H3N8病毒核酸检测结果

, figureFileSmall=null, figureFileBig=null, tableContent=
监测月份（年/月）检测数（份）H3N8阳性标本数(阳性率，%)
2022/04431（2.33）
2022/059818（18.37）
2022/06461（2.17）
2022/08470（0）
2022/09471（2.13）
2022/11471（2.13）
2023/02460（0）
2023/03470（0）
2023/04450（0）
2023/05470（0）
2023/06460（0）
2023/07450（0）
2023/08460（0）
2023/09460（0）
2023/10470（0）
2023/11460（0）
2023/12480（0）
合计83722(2.63)
), ArticleFig(id=1324078128468996699, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, language=EN, label=Table 2, caption=

H3N8 nucleic acid test results from poultry and environmental samples in the epidemic-related live poultry markets

, figureFileSmall=null, figureFileBig=null, tableContent=
样本类型检测数（份）H3N8阳性标本数(阳性率，%)
家禽口咽/泄殖腔拭子2013(1.49)
粪便1442(1.39)
空气1565(3.21)
污水1130（0）
饮水772(2.60)
笼具涂抹837(8.43)
其他涂抹633(4.76)
合计83722(2.63)
), ArticleFig(id=1324078128544494172, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, language=CN, label=表2, caption=

病例涉疫活禽市场家禽及环境样本H3N8病毒核酸检测结果

, figureFileSmall=null, figureFileBig=null, tableContent=
样本类型检测数（份）H3N8阳性标本数(阳性率，%)
家禽口咽/泄殖腔拭子2013(1.49)
粪便1442(1.39)
空气1565(3.21)
污水1130（0）
饮水772(2.60)
笼具涂抹837(8.43)
其他涂抹633(4.76)
合计83722(2.63)
), ArticleFig(id=1324078128624185949, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, language=EN, label=Table 3, caption=

Homology analysis of the H3N8 virus from the epidemic-related live poultry markets and Hunan Province’s first human case of H3N8 infection

, figureFileSmall=null, figureFileBig=null, tableContent=
毒株基因湖南省首例人感染H3N8病例病毒株相似性(%)GenBank登录号
A/environment/Hunan/CS2-0421/2022HAA/Changsha/1000/2022(H3N8)1 743/1 745(99.89)PV490899
NAA/Changsha/1000/2022 (H3N8)1 415/1 418(99.79)PV490900
A/chicken/Hunan/CS20-0525/2022HAA/Changsha/1000/2022 (H3N8)1 743/1 745(99.89)PV490901
NAA/Changsha/1000/2022 (H3N8)1 415/1 418(99.79)PV490902
A/chicken/Hunan/CS51-0927/2022HAA/Changsha/1000/2022 (H3N8)1 741/1 745(99.77)PV495812
NAA/Changsha/1000/2022 (H3N8)1 414/1 418(99.72)PV495813
), ArticleFig(id=1324078128695489118, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, language=CN, label=表3, caption=

病例涉疫活禽市场H3N8病毒与湖南省首例人感染H3N8病例病毒的同源性分析

, figureFileSmall=null, figureFileBig=null, tableContent=
毒株基因湖南省首例人感染H3N8病例病毒株相似性(%)GenBank登录号
A/environment/Hunan/CS2-0421/2022HAA/Changsha/1000/2022(H3N8)1 743/1 745(99.89)PV490899
NAA/Changsha/1000/2022 (H3N8)1 415/1 418(99.79)PV490900
A/chicken/Hunan/CS20-0525/2022HAA/Changsha/1000/2022 (H3N8)1 743/1 745(99.89)PV490901
NAA/Changsha/1000/2022 (H3N8)1 415/1 418(99.79)PV490902
A/chicken/Hunan/CS51-0927/2022HAA/Changsha/1000/2022 (H3N8)1 741/1 745(99.77)PV495812
NAA/Changsha/1000/2022 (H3N8)1 414/1 418(99.72)PV495813
), ArticleFig(id=1324078128770986591, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, language=EN, label=Table 4, caption=

Comparative analysis of key sites in the HA and NA proteins of viruses isolated from the epidemic-related live poultry markets and Hunan Province’s first human case of H3N8 infection

, figureFileSmall=null, figureFileBig=null, tableContent=
毒株名称HA蛋白受体结合位点HA蛋白裂解位点NA蛋白耐药位点NA蛋白缺失位点
242～244a340～348aH273Y aR291K a69～73
A/environment/Hunan/CS2-0421/2022(H3N8)QSGPEKQTRGLFHR否
A/chicken/Hunan/CS20-0525/2022(H3N8)QSGPEKQTRGLFHR否
A/chicken/Hunan/CS51-0927/2022(H3N8)QSGPEKQTRGLFHR否
A/Changsha/1000/2022(H3N8)QSGPEKQTRGLFHR否
), ArticleFig(id=1324078128854872672, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1324078120709534234, language=CN, label=表4, caption=

病例涉疫活禽市场H3N8病毒与湖南省首例人感染H3N8病例病毒株HA和NA蛋白关键位点比对分析

, figureFileSmall=null, figureFileBig=null, tableContent=
毒株名称HA蛋白受体结合位点HA蛋白裂解位点NA蛋白耐药位点NA蛋白缺失位点
242～244a340～348aH273Y aR291K a69～73
A/environment/Hunan/CS2-0421/2022(H3N8)QSGPEKQTRGLFHR否
A/chicken/Hunan/CS20-0525/2022(H3N8)QSGPEKQTRGLFHR否
A/chicken/Hunan/CS51-0927/2022(H3N8)QSGPEKQTRGLFHR否
A/Changsha/1000/2022(H3N8)QSGPEKQTRGLFHR否
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2022—2023年长沙市病例涉疫活禽市场中H3N8禽流感病毒分子监测
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张如胜 1 , 扶会媛 1 , 裴瑞青 1 , 黄政 1 , 罗璨 1 , 陈静芳 1 , 陈姗 1 , 陈腊梅 2
现代预防医学 | 疾病预防控制 2026,53(3): 533-538
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现代预防医学 |疾病预防控制 2026 , 53 (3) : 533 -538
2022—2023年长沙市病例涉疫活禽市场中H3N8禽流感病毒分子监测
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张如胜（1977-），男，硕士，主任技师，研究方向：新发传染病的分子流行病学研究

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张如胜（1977-），男，硕士，主任技师，研究方向：新发传染病的分子流行病学研究

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张如胜1, 扶会媛1, 裴瑞青1, 黄政1, 罗璨1, 陈静芳1, 陈姗1, 陈腊梅2
作者信息
  • 1.长沙市疾病预防控制中心(长沙市卫生综合监督执法局)，湖南 长沙 410007
  • 2.开福区疾病预防控制中心(开福区卫生综合监督执法局)，湖南 长沙 410007
通讯作者:
陈腊梅，E-mail:
作者简介:

张如胜（1977-），男，硕士，主任技师，研究方向：新发传染病的分子流行病学研究

Molecular surveillance of H3N8 avian influenza virus in live poultry markets associated with human cases in Changsha City, 2022—2023
Ru-sheng ZHANG1, Hui-yuan FU1, Rui-qing PEI1, Zheng HUANG1, Can LUO1, Jing-fang CHEN1, Shan CHEN1, La-mei CHEN2
Affiliations
  • Changsha Center for Disease Control and Prevention (Changsha Municipal Health Comprehensive Supervision and Law Enforcement Bureau), Changsha, Hunan 410007, China
出版时间: 2026-02-10 doi: 10.20043/j.cnki.MPM.202508170
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摘要
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目的

分析湖南省首例人感染H3N8亚型禽流感病毒（avian influenza virus, AIV）（简称H3N8病毒）病例涉疫活禽市场中H3N8病毒污染状况及其分子进化特征，为禽流感疫情防控提供依据。

方法

于2022年4月—2023年12月（共17个月）对湖南省首例人感染H3N8病例的涉疫活禽市场进行H3N8病毒监测，采集家禽口咽/泄殖腔拭子、粪便、空气、污水、饮水、笼具涂抹物及其他物体表面涂抹样本共837份。采用Real-Time PCR技术检测样本中H3N8病毒核酸，对阳性样本进行高通量核苷酸测序，对获得的病毒血凝素（hemagglutinin, HA）和神经氨酸酶（neuraminidase, NA）基因序列进行BLAST分析，利用BioEdit和Mega X软件分别进行氨基酸序列比对及系统进化树构建。核酸检测结果统计学分析方法采用Fisher精确检验。

结果

样本总体H3N8病毒核酸阳性率为2.63% (22/837)。病例发生当月（2022年5月）阳性率最高，为18.37%，显著高于其他16个月份（χ2=49.159, P<0.001）。6类样本中检出病毒核酸，其中笼具涂抹样本阳性率最高，为8.43%（χ2=14.437, P=0.011）。测序获得3份来源于家禽饮水和鸡口咽/泄殖腔拭子样本的H3N8病毒，其HA、NA基因与人源株（A/China/CSKFQ-22-5/2022）核苷酸相似性最高（99.71～99.89%）。HA蛋白裂解位点（第340～348位氨基酸）为PEKQTRGLF，受体结合位点（第242、244位氨基酸）为Q、G，呈现低致病性及禽类受体结合特征。NA蛋白未发现神经氨酸酶抑制剂耐药位点（H273Y、R291K）突变。系统进化分析显示，3株病毒的HA、NA基因均属欧亚分支，并与国内人、鸡及环境来源的H3N8病毒共同形成一个独立进化分支。

结论

活禽市场H3N8病毒污染程度与人感染病例的发生存在一定相关性，应密切监测活禽市场中的H3N8病毒污染状况。

活禽市场  /  环境  /  禽流感病毒  /  H3N8亚型  /  基因
Objective

To characterize H3N8 virus contamination and molecular evolution in poultry and environmental samples from the live poultry markets associated with Hunan Province’s first human H3N8 avian influenza virus (AIV) case, providing evidence for avian influenza epidemic prevention and control.

Methods

Active surveillance for H3N8 avian influenza virus was implemented over 17 months (April 2022—December 2023) in live poultry markets associated with Hunan Province’s first human case. We collected 837 samples, including poultry oropharyngeal/cloacal swabs, feces, air, sewage, drinking water, cage swabs, and other surface swabs. Samples were screened for H3N8 nucleic acid using real-time PCR. Positive specimens underwent high-throughput sequencing. Hemagglutinin (HA) and neuraminidase (NA) gene sequences were analyzed by BLAST, with amino acid alignment and phylogenetic tree construction performed using BioEdit and MEGA-X, respectively.The statistical analysis method for nucleic acid test results was Fisher’s exact test.

Results

The overall H3N8 nucleic acid positivity rate was 2.63% (22/837). The highest positivity rate (18.37%) occurred in the month of the human case (May 2022),significantly higher than in the other 16 months (χ2=49.159, P<0.001). Viral nucleic acid was detected in six sample types, with cage swabs showing the highest positivity rate (8.43%, χ2=14.437, P=0.011). Three H3N8 viruses were successfully sequenced from poultry drinking water and chicken oropharyngeal/cloacal swabs. Their HA and NA genes exhibited the highest nucleotide similarity (99.71-99.89%) to the human-origin strain (A/China/CSKFQ-22-5/2022). The HA protein cleavage site (residues 340-348) was PEKQTRGLF, and the receptor-binding site residues (242 and 244) were Q and G, indicating low pathogenicity and avian receptor-binding specificity. No neuraminidase inhibitor resistance mutations (H273Y, R291K) were detected in the NA protein. Phylogenetic analysis revealed that the HA and NA genes of all three viruses belonged to the Eurasian lineage and formed a distinct subclade with H3N8 viruses of human, chicken, and environmental origin from China.

Conclusion

The level of H3N8 virus contamination in live poultry markets shows an association with human infection cases. Enhanced surveillance of H3N8 virus contamination in these markets is warranted.

Live poultry market  /  Environment  /  Avian influenza virus  /  H3N8 subtype  /  Gene
张如胜, 扶会媛, 裴瑞青, 黄政, 罗璨, 陈静芳, 陈姗, 陈腊梅. 2022—2023年长沙市病例涉疫活禽市场中H3N8禽流感病毒分子监测. 现代预防医学, 2026 , 53 (3) : 533 -538 . DOI: 10.20043/j.cnki.MPM.202508170
Ru-sheng ZHANG, Hui-yuan FU, Rui-qing PEI, Zheng HUANG, Can LUO, Jing-fang CHEN, Shan CHEN, La-mei CHEN. Molecular surveillance of H3N8 avian influenza virus in live poultry markets associated with human cases in Changsha City, 2022—2023[J]. Modern Preventive Medicine, 2026 , 53 (3) : 533 -538 . DOI: 10.20043/j.cnki.MPM.202508170
正文
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禽流感病毒（avian influenza virus，AIV）根据表面糖蛋白血凝素（hemagglutinin, HA）和神经氨酸酶（neuraminidase, NA）可分为18个HA亚型（H1～H18）和11个NA亚型（N1～N11），其中H3亚型AIV通常在野生禽类或家鸭中流行，较少导致鸡群暴发疫情，然而，自2021年12月起，我国广东、河南等地的鸡群中出现了新型重配H3N8亚型AIV（简称H3N8病毒）疫情，研究显示该病毒可在鸡群中快速传播，感染特征包括眼睑急性肿胀、出血并伴有呼吸道症状等[1-2]。2022年4月，H3N8病毒首次发生跨物种传播，河南省驻马店市报告了全球首例人感染H3N8病毒引发的重症肺炎病例[3]。次月（2022年5月），湖南省长沙市报告了该省首例人感染H3N8病毒病例，流行病学调查表明其感染来源为涉疫活禽市场暴露[4]。值得注意的是，活禽市场环境已被确认为H5N6、H7N9等人感染AIV的重要来源[5-6]。
既往研究也显示长沙市辖区活禽市场存在较普遍的AIV环境污染[7-9]。病毒的致病能力、受体结合特性及耐药性与其基因特征密切相关。值得关注的是，全球首例与湖南省首例人感染H3N8病例的临床症状存在差异，前者表现为重症肺炎，后者表现为流感样症状。研究指出，这种差异与两例病例病毒分离株基因编码的特定氨基酸（amino acid, AA）位点变异有关[10-11]。湖南省首例人感染H3N8病例疫情发生后，其涉疫活禽市场家禽及环境的H3N8病毒污染状况与分子进化特征，成为疾病防控的关键科学问题。因此，本研究于2022年4月—2023年12月，采集涉疫活禽市场家禽及环境样本进行H3N8病毒核酸检测和进化分析，为H3N8病毒疫情防控措施的制定提供依据。
1　材料与方法
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1.1　监测点
2022年5月，长沙市疾病预防控制中心检出湖南省首例人感染H3N8禽流感病例，选择该病例暴露过的长沙市开福区某农贸市场，以及为其供货的长沙县某活禽批发市场作为涉疫活禽市场监测点[4,11]。
1.2　家禽及环境样本采集
2022年5月—2023年12月，对病例涉疫活禽市场开展了家禽及环境样本采集工作。此外，还对病例发生前一个月（2022年4月）的涉疫活禽市场日常监测留存样本进行了回顾性H3N8禽流感监测，总监测时间为17个月。采样点包括病例暴露的某农贸市场（仅在2022年5月采样一次）以及为其供货的某活禽批发市场（每1～2个月采样一次）。每次采样时，按摊位顺序选取4～5家店铺，分别采集家禽口咽/泄殖腔拭子、粪便、空气、污水、家禽饮水、笼具和其他涂抹等多种类型样本，每类样本采集1～2份。平均每次采样约50份样本。使用病毒采样保存液（北京友康生物科技股份有限公司）共采集家禽及环境样本837份，其中家禽样本包括家禽口咽/泄殖腔拭子201份和粪便144份；环境样本包括空气156份、污水113份、饮水77份、笼具涂抹83份及其他涂抹样本63份。各样本采集方法参考文献[12-13]。
1.3　AIV核酸检测
家禽及环境样本使用自动核酸提取仪及配套提取试剂（西安天隆科技有限公司）进行核酸提取，然后采用禽流感病毒H3N8亚型核酸检测试剂盒（双重荧光 PCR 法）（深圳市梓健生物科技有限公司）及ABI 7500型Real-time PCR仪（美国Thermo Fisher Scientific公司）对提取的样本核酸进行H3N8病毒核酸检测。
1.4　AIV核酸序列测定
采用AIV通用引物（Uni-12/Inf1：5’-GGGGGGAGCAAAAGCAGG-3’，Uni-12/Inf3：5’-GGGGGGAGCGAAAGCAGG-3’Uni-13/Inf1：5’-CGGGTTATTAGTAGAAACAAGG-3’）illumina Miseq测序仪及双端150 bp测序试剂盒（美国illumina公司）对H3N8病毒阳性样本核酸进行序列测定，核酸起始量范围为1～500 ng DNA，建库采用DNA prep文库构建试剂盒（美国illumina公司）。下机数据先用Fastqc、Cutadapt和Bowite2软件进行数据质量分析、去接头和reads比对，测序数据经SPAdes-3.13.0软件拼接后提交至基因序列数据库（Genetic Sequence Database，Genbank），在线基本局部相似性搜索工具（basic local alignment search tool, BLAST）进行核苷酸相似性分析（http：//BLAST.ncbi.nlm.nih.gov/BLAST.cgi）。
1.5　H3N8病毒基因AA关键位点分析
从全球流感数据共享倡议（Global Initiative on Sharing All Influenza Data，GISAID）和美国国家生物技术信息中心（National Center for Biotechnology Information,NCBI）下载H3N8、H5N1、H5N6、H6N6、H7N9、H9N2、H10N8及H11N3病毒代表株基因编码AA序列，利用BioEdit软件进行基因编码关键AA位点比对。从GISAID下载2022—2024年部分H3N8病毒 HA和NA基因核苷酸序列，首先利用MEGA X软件进行DNA序列ALIGN Clustal W比对，对首尾长度不一致的核苷酸序列进行删除，然后利用MODELS程序Find Best DNA/Protein Models（ML）预测分析模型，根据模型预测结果，构建HA和NA基因进化树分别选择Tamura三参数模型(Tamura 3-parameter Model，T92）+Gamma分布（G）和T92+G+不变位点比例（I），方法选择最大似然法（maximum likelihood），bootstrap重复次数设为1 000次。
1.6　H3N8病毒HA、NA基因进化分析
1.7　统计分析
家禽及环境样本H3N8病毒核酸检测结果应用SPSS 24.0软件进行统计学分析，不同监测时间及不同样本类型阳性率间的比较均采用Fisher精确检验，检验水准α=0.05。
2　结果
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2.1　H3N8病毒不同时间监测情况
病例涉疫活禽市场家禽及环境样本的H3N8病毒总体阳性率为2.63%，病例发生当月（2022年5月）H3N8病毒阳性率最高，为18.37%，显著高于其他监测月份（0～2.33%），2023年全年未检出H3N8病毒阳性，具体结果见表1。经Fisher精确检验，不同监测时间的H3N8病毒阳性率差异具有统计学意义（χ2=49.159, P<0.001）。
2.2　H3N8病毒不同样本监测情况
家禽口咽/泄殖腔拭子、粪便、空气、污水、饮水、笼具涂抹和其他涂抹样本的H3N8阳性率分别为1.49%（3/201）、1.39%（2/144）、3.21%（5/156）、0（0/113）、2.60%（2/77）、8.43%（7/83）和4.76%（3/63），具体结果见表2。统计学分析显示笼具涂抹H3N8病毒阳性率显著高于其余样本类型，不同样本类型间的阳性率差异具有统计学意义（χ2=14.437, P=0.011）。
2.3　H3N8病毒核苷酸序列同源性分析
22份H3N8病毒核酸阳性样本进行高通量核苷酸测序和BLAST比对，获得3份分别来源于家禽饮水（1份）和鸡口咽/泄殖腔拭子（2份）的H3N8病毒，3份样本来源的H3N8病毒HA及NA基因序列测序深度符合质量控制要求，且均与湖南省首例人感染H3N8病毒（A/Changsha/1000/2022）核苷酸相似性最高，相关 H3N8病毒的核苷酸序列已上传至 GenBank，登录号分别为 PV490899-PV490902、PV495812 和 PV495813。具体比对结果见表3。
2.4　H3N8病毒HA和NA基因编码AA关键位点比对分析
本研究3份样本来源的H3N8 病毒HA和NA基因关键位点与湖南省首例人感染H3N8病毒（A/Changsha/1000/2022）相同，其中HA蛋白裂解位点（第340～348位AA）均为PEKQTRGLF，HA蛋白受体结合位点（receptor binding site，RBS）第242和244位（对应H3流感病毒第226和228位）AA均为Q和G，NA蛋白第273和291位AA均为为H和R，且第69～73位点未发生缺失，具体比对结果见表4。
2.5　H3N8病毒HA、NA基因进化树构建
结果显示，H3N8病毒HA和NA基因进化树均形成欧亚和北美两大分支，本研究中3份样本来源的H3N8病毒与湖南省首例人感染H3N8病毒（A/Changsha/ 1000/2022）及湖南鸡源H3N8病毒（A/chicken/ Hunan/S2342/2022）亲缘关系近，属于欧亚大分支，且已与其他H3N8病毒中国分离株进化形成一个独立的亚分支。北美大分支则主要由源自美国、加拿大等北美地区的野鸟及家禽 H3N8 病毒组成。见图1。
3　讨论
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既往研究显示活禽市场家鸭存在H3亚型AIV流行[2,14]，以H3N2病毒为主，H3N8病毒较为少见[15]。本研究显示，2022—2023年湖南省首例人感染H3N8病例涉疫活禽市场家禽及环境样本的H3N8病毒总体阳性率为2.63%。值得注意的是，病例发生当月（2022年5月）的病毒阳性率最高，达到18.37%；而病例发生前一个月（2022年4月）及随后数月（2022年6、9和11月）的阳性率均维持在较低水平（约2.00%）。需要指出的是，病例发生后实施的强化消杀措施是导致阳性率下降的因素之一。此外，2023年全年监测中未检出H3N8病毒核酸阳性，同期辖区内也无新发人感染H3N8病例报告。以上结果表明，该病例的出现与涉疫活禽市场中H3N8病毒污染程度的升高存在一定关联。类似地，其他研究也表明活禽市场环境中污染H7N9 病毒后，则暴露于活禽市场的人群存在感染风险[6]。因此，建议相关部门加强H3N8 病毒在禽类及环境中的监测力度，以便及时预警人感染风险。
研究表明，鸡感染H3N8病毒后，病毒可在多组织脏器中广泛分布，其中在呼吸系统和眼球附近的哈德氏腺病毒含量最高，病毒主要经口咽和泄殖腔排出[1]，造成环境污染及易感鸡群感染。本研究在病例涉疫活禽市场家禽及环境样本采集过程中，观察到活禽销售店铺存在笼具/圈养空间狭小、家禽密度高、通风不良等问题。这种环境一旦出现感染禽只，极易导致病毒在待售禽群间传播并污染环境。本次病例涉疫活禽市场监测数据显示，H3N8病毒在6种监测样本类型中检测到H3N8病毒，其中笼具涂抹样本阳性率最高，为8.43%，特别是在活禽市场空气中监测到H3N8病毒，结合湖南省首例人感染H3N8病例的流行病学调查（暴露于活禽市场但未直接接触活禽）以及文献报道的 H3N8 病毒可在雪貂间经空气传播的证据[4,10]，提示湖南省首例人感染H3N8病例存在经含病毒气溶胶暴露感染的可能性。因此，建议相关部门在加强活禽市场禽类环境清洁消毒的同时，应着重改善场所的通风条件，以降低此类传播风险。
AIV通常优先结合禽类α2,3-Gal唾液酸受体，若其结合人类α2,6-Gal唾液酸受体的能力增强，则表明对哺乳动物的适应性提升。研究表明，H3N2和H3N8病毒具备同时结合禽类和人类双重受体的能力[16-17]。本研究测序获得1份来源于家禽饮水和2份来源于鸡口咽/泄殖腔拭子的H3N8病毒，其HA蛋白裂解位点和受体结合位点分析显示低致病性特征，并主要结合禽类α2,3-Gal受体。NA基因耐药位点分析提示对神经氨酸酶抑制剂药物（奥司他韦等）敏感，与湖南省首例人源H3N8病毒株分子特征相似[18]，但与全球首例人感染H3N8病例病毒株具备哺乳动物适应及致病能力增强的分子特征存在差异[11]，提示目前国内流行的H3N8病毒具备潜在的大流行风险。
研究表明，H3亚型AIV在国内活禽市场的家鸭中广泛流行并持续发生重配[19]，HA和NA基因可分为欧亚分支和北美分支。H3N8病毒的来源复杂，其HA基因可能源自欧亚地区家禽中流行的H3亚型AIV，而NA基因可能来自东亚、北美和欧洲。欧亚分支在国内普遍存在，而北美分支主要分布于中国东部地区[20]。本研究构建的2022—2024年H3N8病毒HA和NA基因进化树显示：包括本研究3份样本来源的H3N8病毒在内的H3N8病毒中国分离株已形成一个独立的亚分支。此外，NA基因的欧亚分支还包含来自加拿大、美国等北美地区环境和野鸟H3N8病毒株，进一步证实了新型重配H3N8病毒由欧亚分支的H3基因、北美分支的N8基因与G57基因型H9N2病毒的内部基因三源重配组成的研究结论[1]。进化分析还表明，3份样本来源的H3N8病毒与湖南省首例人感染H3N8病毒（A/Changsha/1000/2022）及湖南鸡源H3N8病毒（A/chicken/Hunan/S2342/2022）亲缘关系近，进一步证实该人感染病例的病毒来源于涉疫活禽市场的鸡源 H3N8 病毒。
既往研究指出，H9N2 亚型AIV在鸡群中的流行及其在活禽市场环境中的污染最为普遍[8,21]。这一现象主要归因于当前鸡H9N2病毒灭活疫苗的局限性：虽然能够减轻感染临床症状，但无法有效阻断病毒在鸡上呼吸道的定植与排毒，导致免疫鸡群仍可持续携带并传播病毒[22]。这也是H9N2病毒为新型重配H7N9、H3N2、H10N8、H5N6及H3N8[11,23]亚型AIV提供内部基因来源的重要原因，因此加强新型H9N2病毒疫苗研发是控制家禽H9N2病毒疫情的关键措施，阻断其作为新型重配AIV的内部基因来源，是预防控制人感染H3N8等新亚型重配病毒疫情的根本方法[1]。
综上所述，本研究提示病例涉疫活禽市场中H3N8病毒的污染水平与人感染病例的发生存在一定相关性。尽管目前人感染H3N8病例仍为散发，但考虑到H3亚型AIV的流感大流行潜力，H3N8病毒表现出α 2,3-Gal与α2,6-Gal唾液酸双受体的结合特性以及人群普遍缺乏对H3N8病毒感染的特异性免疫力[24]，因此建议相关部门加强H3N8病毒监测，建立“禽-环境-人”传播链的早期预警体系，防范流感大流行。
基金
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  • 湖南省自然科学基金(2023JJ60399; 2023JJ60070)
  • 湖南省卫生健康高层次人才重大科研专项(R2023169)
  • 湖南省卫生健康委科研项目(202203094936)
  • 长沙市卫健委科研项目(KJ-B2023099)
  • 长沙市自然科学基金项目(kq2403183)
文献
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参考文献 引证文献
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2026年第53卷第3期
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doi: 10.20043/j.cnki.MPM.202508170
  • 接收时间：2025-08-13
  • 首发时间：2026-11-02
  • 出版时间：2026-02-10
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  • 收稿日期：2025-08-13
基金
湖南省自然科学基金(2023JJ60399; 2023JJ60070)
湖南省卫生健康高层次人才重大科研专项(R2023169)
湖南省卫生健康委科研项目(202203094936)
长沙市卫健委科研项目(KJ-B2023099)
长沙市自然科学基金项目(kq2403183)
作者信息
    1.长沙市疾病预防控制中心(长沙市卫生综合监督执法局)，湖南 长沙 410007
    2.开福区疾病预防控制中心(开福区卫生综合监督执法局)，湖南 长沙 410007

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陈腊梅，E-mail:
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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