摘要
目的对活禽市场家禽中分离的H5N6禽流感病毒(avian influenza virus,AIVs)进行遗传进化和分子特征分析。方法2018年12月于乌鲁木齐市活禽市场采集家禽双腔拭子和环境样本,通过接种鸡胚、血凝试验和RT-PCR等方法分离鉴定AIVs,以甲型流感病毒通用引物扩增病毒全基因组并测序,利用BLAST、Clustal W、MEGA-X和DNAStar等软件进行序列比对、系统进化和分子特征分析。结果从家禽样品中分离到6株H5N6亚型AIVs,其各基因之间的一致性较高(99.4%~100.0%),为同一来源。BLAST分析显示除NP基因与苏州家禽中分离的H5N6毒株一致性最高(99.7%),其余7个基因片段均与2017—2018年广东环境中分离的H5N6毒株一致性最高,在99.0%~100.0%之间。遗传进化分析显示分离株的HA属于Clade2.3.4.4.C分支,HA、NA、PB1、PA、NP、MP这6个基因均与2018年自华东地区水貂中分离的H5N6病毒株聚在同一分支,PB2和NS均与2017—2018年于广东环境中分离的H5N6病毒株聚在一起。分离株HA的裂解位点含多个连续的碱性氨基酸,为高致病性AIV,HA的S137A和T160A突变可增强病毒与人源受体SAα2,6-Gal的结合。另外,NA茎部缺失、PB2的L89V、G309D、R477G、I495V、I504V、D391E和A661E以及NS1的P42S、D92E和80~84位氨基酸缺失等突变均可增强病毒对哺乳动物的毒力和致病力。结论分离的6株高致病性H5N6 AIVs与2017—2018年分离自华东地区水貂和广东环境的H5N6 AIVs的遗传关系较近,存在的多个突变可增强病毒对哺乳动物的致病性,存在较大的公共卫生风险。
Objective To analyze phylogenetic structure and molecular characteristics of H5N6 avian influenza virus(AIVs)isolated from live poultry market(LPM).Methods Oropharyngeal and cloacal swabs from poultry,and environmental samples were collected from LPM in Urumqi in December 2018,AIVs were isolated and identified by inoculation of chicken embryo,hemagglutination test and RT-PCR,the viral whole genome was amplified with the universal primers of influenza A virus,and then sequenced,pairwise sequence alignments,phylogenetic and molecular characteristics analysis were performed by BLAST,Clustal W,MEGA-X and DNAStar software.Results Six strains of H5N6 AIVs were isolated from poultry samples,the identity between the viral genes was high(99.4%-100.0%),so the isolates were the same source.BLAST analysis revealed that the viral NP sequence had the highest identity(99.7%)with H5N6 AIVs isolated from poultry in Suzhou,while the sequence of the remaining 7 viral genes had the highest identity(99.0%-100.0%)with H5N6 AIVs isolated from environment in Guangdong during 2017 to 2018.Phylogenetic analysis showed that the viral HA belonged to Clade 2.3.4.4C,and the viral HA,NA,PB1,PA,NP,and MP were all clustered together with H5N6 AIVs isolated from mink in Eastern China in 2018,while the PB2 and NS were clustered together with H5N6 AIVs isolated from environment in Guangdong from 2017 to 2018.The HA cleavage site contained multiple basic amino acid residues,which was highly pathogenic AIVs(HPAIVs).S137A and T160A mutations of HA could increase binding to human-type receptor SAα2,6-Gal.Additionally,the viral multiple mutations,including 59-69 deletion in NA,the L89V,G309D,R477G,I495V,I504V,D391E,and A661E in PB2,as well as the P42S,D92E,and 80-84 deletion in NS1,could enhance the viral virulence and pathogenicity to mammals.Conclusions The 6 strains of H5N6 HPAIVs isolated from LPM have relatively close genetic relationship with H5N6 AIVs isolated from mink in Eastern China and environment in Guangdong during 2017 to 2018,the viral multiple mutations could increase its pathogenicity to mammals,which could pose a potential risk to public health.
作者
常娜娜
张倩
张成
杜菲
阿木提喀日·马木提
毕玉海
马正海
Chang Nana;Zhang Qian;Zhang Cheng;Du Fei;Amutikari·Mamuti;Bi Yuhai;Ma Zhenghai(Xinjiang Key Laboratory of Biological Resources and Genetic Engineering,College of Life Science and Technology,Xinjiang University,Urumqi 830046,China;Center for Influenza Research and Early-Warning,Key Laboratory of Pathogenic Microbiology and Immunology,Institute of Microbiology,Chinese Academy of Science,Beijing 100101,China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2021年第4期254-261,共8页
Chinese Journal of Microbiology and Immunology
基金
新疆维吾尔自治区区域协同创新专项(上海合作组织科技伙伴计划)(2017E01022)
国家科技重大专项(2018ZX10101004)。
关键词
禽流感病毒
H5N6
活禽市场
遗传进化
Avian influenza virus
H5N6
Live poultry market
Genetic evolution
作者简介
通信作者:马正海,Email:mzhxju@126.com,电话:0991-8582554。
