2021—2023年福建省活禽市场禽流感病毒监测和H9N2亚型序列分析  

Surveillance of avian influenza virus and sequence analysis of H9N2 subtype in live poultry markets in Fujian province,2021-2023

作　　者：吴晶晶 张雅婷 鄢育青 陈宏彬[1] 林琦[1] 翁育伟[1,2] Wu Jingjing;Zhang Yating;Yan Yuqing;Chen Hongbin;Lin Qi;Weng Yuwei(Fujian Center for Disease Control and Prevention,Fujian Provincial Key Laboratory of Zoonosis Research,Fuzhou 350012,China;The School of Public Health,Fujian Medical University,Fuzhou 350122,China)

机构地区：[1]福建省疾病预防控制中心,福建省人兽共患病研究重点实验室,福州350012 [2]福建医科大学公共卫生学院,福州350122

出　　处：《中华实验和临床病毒学杂志》2025年第1期41-50,共10页Chinese Journal of Experimental and Clinical Virology

基　　金：福建省自然科学基金项目(2021J01351);福建省卫生健康中青年科研重大项目(2021ZQNZD006);福建省卫健委青年科研课题(2022QNA059)。

摘　　要：目的分析2021—2023年福建省活禽市场禽流感病毒流行特点及H9N2亚型禽流感病毒分子和遗传特征,为禽流感防控、预测工作提供参考。方法在福建省5个地市的活禽市场监测点进行6种不同类型环境标本的采样,采用荧光定量PCR方法对环境标本进行A型、H5、H7、H9亚型核酸检测,并采用χ^(2)检验进行统计分析。选取部分Ct值<31的标本,对血凝素(hemagglutinin,HA)和神经氨酸酶(neuraminidase,NA)基因进行靶向扩增和二代测序,下载参考序列,使用生物学信息软件进行氨基酸突变和遗传进化分析。结果2021—2023年共采集5个监测点1853份标本,A型流感病毒阳性率为50.94%(944份),其中H9亚型684份、H5亚型23份、H7亚型1份、H5+H9亚型28份、H7+H9亚型1份、A型未分型标本207份。不同地市A型流感病毒阳性检出率存在显著差异(χ^(2)=461.82,P<0.001);不同年份和不同季度A型流感病毒阳性率的差异具有统计学意义(χ^(2)=12.26,P=0.002;χ^(2)=30.12,P<0.001),第一季度(56.39%)和第三季度(55.34%)的阳性率较高;不同类型标本A型阳性率的差异具有统计学意义(χ^(2)=23.05,P<0.001),笼具表面擦拭标本的阳性率最高(56.09%)。共完成24份H9N2亚型阳性样本的HA和NA基因测序。与NCBI数据库序列比对,HA基因核苷酸序列最高一致性在97.03%~99.87%,NA基因核苷酸序列最高一致性在97.50%~99.78%;23株毒株的HA和NA基因都归属于欧亚谱系Y280-like进化分支G57基因型,另有1株归属于欧亚谱系Y439-like进化分支;24株毒株的裂解位点均为低致病性禽流感病毒分子特征,大部分毒株受体结合位点为禽源和人类双受体结合特征。结论2021—2023年福建省活禽市场存在较高阳性率的禽流感病毒污染,第一季度和第三季度为重点防控季度,以H9亚型为主要亚型,大部分H9N2亚型HA和NA基因归属于Y280-like G57基因型,伴随少量Y439-like进化分支,提示基因重组的可能,存在人类感染的�ObjectiveTo analyze the epidemiologic characteristics of avian influenza virus and the molecular characteristics and genetic evolution of H9N2 subtype avian influenza viruses in the live poultry markets in Fujian province in 2021-2023.MethodsSix types of specimens were collected from five cities in Fujian province.The specimens were subjected to nucleic acid detection for influenza A viruses,subtypes H5,H7,and H9 by fluorescence quantitative PCR,and the results were analyzed statistically withχ^(2) test.Specimens with Ct value less than 31 were screened for targeted amplification and next-generation sequencing of the hemagglutinin(HA)and neuraminidase(NA)genes.Reference sequences were downloaded from the databases,and the characteristics of molecular variation and genetic evolution were analyzed by using bioinformatics softwares.ResultsFrom 2021 to 2023,a total of 1853 specimens were collected from five cities,with a positive rate of 50.94%for influenza A viruses,including 684 specimens of H9 subtype,23 specimens of H5 subtype,1 specimen of H7 subtype,28 specimens of H5 and H9 subtype,1 specimen of H7 and H9 subtype,and 207 specimens of unclassified A-type.There were significant differences in the positive detection rates of influenza A viruses in different cities(χ^(2)=461.82,P<0.001).Statistically significant differences in influenza A virus positivity rates across years and quarters(χ^(2)=12.26,P=0.002;χ^(2)=30.12,P<0.001),with higher rates of 56.39%and 55.34%in the first and third quarters,respectively.And the differences in the positive rates of influenza A viruses in different types of specimens were statistically significant(χ^(2)=23.05,P<0.001),with specimens on the cage surface having a positivity rate of 56.09%,which was the highest of all types of specimens.A total of 24 strains of HA and NA genes of H9N2 subtype were fully sequenced.Compared with the sequence of NCBI database,the highest identity of HA gene nucleotide sequences ranged from 97.03%to 99.87%,and the highest identity of NA gene nucle

关 键 词：禽流感病毒 H9N2亚型 监测 遗传进化 变异 

分 类 号：S85[农业科学—兽医学]