中国南方三省H10亚型禽流感病毒基因特征分析

doi:10.6040/j.issn.1671-7554.0.2021.0220

摘要/Abstract

摘要：

目的: 分析2014~2019年我国南方三省H10亚型禽流感病毒的基因特征。方法: 采集2014~2019年江西、湖南和广西3个省份活禽市场环境样本共计30231份，采用实时荧光定量PCR对上述环境样本进行甲型禽流感病毒筛查，阳性样本进一步做病毒分离，若病毒分离结果为阳性，则对其进行基因深度测序。利用CLC、CD-HIT、MAFFT v7.037和MEGA v6.06软件分析流感病毒序列。结果: 三省禽流感强化监测共分离到11株H10亚型禽流感病毒。通过全基因组测序和系统进化分析发现，除3个N8基因位于北美分支，其他基因均位于欧亚分支。JX24472与人H10N8亚型禽流感病毒HA基因片段亲源关系较近。江西分离的5株H10N3亚型禽流感病毒内部基因片段较为分散，内部基因片段来源呈多样性。病毒蛋白位点分析结果显示部分潜在的与受体结合位点、小鼠致病性以及在哺乳动物中的传播性有关的氨基酸位点发生突变。结论: 本研究分离的11株H10亚型禽流感病毒与人H10N8亚型禽流感病毒基因处于不同分支(JX24472除外)，但是氨基酸位点分析提示这些病毒具有潜在使人或者禽感染并致病的可能性，应持续对其加强监测，为潜在的流感大流行做准备。

关键词: 禽流感病毒, H10N3, H10N8, 进化分析, 活禽市场, 监测

Abstract:

Objective: To explore the genetic characteristics of avian influenza virus (AIVs) between 2014 and 2019 in three southern provinces of China. Methods: A total of 30 231 environment samples isolated from Jiangxi, Hunan and Guangxi provinces from 2014 to 2019 were tested for flu A with real-time RT-PCR. If the testing results were positive, the samples were propagated in SPF chicken embryonated eggs. All positive AIVs isolations were sequenced. CLC, CD-HIT, MAFFT v7.037 and MEGA v6.06 software were used to analyze the influenza sequences. Results: Altogether 11 H10 subtype AIVs were isolated. Whole genome sequencing and phylogenetic analysis revealed that all genes were classified into Eurasian lineage, except 3 N8 genes in the North American lineage. JX24472 had close relationship with human infection H10N8 AIVs in its HA segment. The internal genome of 5 H10N3 subtype AIVs isolated from Jiangxi Province suggested their genome were diverse and complex. Molecular characteristic analysis of H10 subtype AIVs revealed that several mutations in key amino acid sites were associated with receptor binding sites, pathogenicity in mice and transmissibility in mammals. Conclusion: Phylogenetic analyses of the 11 H10 subtype AIVs isolated in this research suggested that they were in different clade with human infected H10N8 AIVs, except JX24472. Whereas the amino acid characterization analysis suggested that these H10 subtype viruses had the potential to infect human or poultry to make them sick. Therefore, it is necessary to strengthen the continuous surveillance of H10 subtype AIVs and prepare for the next potential pandemic.

Key words: Avian influenza virus, H10N3, H10N8, Phylogenetic analysis, Live poultry market, Surveillance

中图分类号:

R373.1

曾晓旭,张恒,焦铭,陈涛,李希妍,黄维娟,杨静,王大燕. 中国南方三省H10亚型禽流感病毒基因特征分析[J]. 山东大学学报 (医学版), 2021, 59(5): 52-59.

Xiaoxu ZENG,Heng ZHANG,Ming JIAO,Tao CHEN,Xiyan LI,Weijuan HUANG,Jing YANG,Dayan WANG. Genetic characteristics of H10 subtype avian influenza virus in three southern provinces of China[J]. Journal of Shandong University (Health Sciences), 2021, 59(5): 52-59.

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参考文献 30

1	Chen HY , Yuan H , Gao RB , et al. Clinical and epidemiological characteristics of a fatal case of avian influenza A H10N8 virus infection: a descriptive study[J]. Lancet, 2014, 383 (9918): 714- 721. doi: 10.1016/S0140-6736(14)60111-2
2	杨树, 夏文, 刘明斌, 等. 2014年江西省南昌市一例人感染H10N8禽流感病例流行病学调查[J]. 疾病监测, 2015, 30 (5): 428- 430.
	YANG Shu , XIA Wen , LIU Mingbin , et al. Epidemiological survey of a case of human infection with avian influenza A (H10N8) virus in Nanchang, Jiangxi, 2014[J]. Disease Surveillance, 2015, 30 (5): 428- 430.
3	傅伟杰, 胡茂红, 刘晓青, 等. 江西省3例H10N8禽流感病毒感染者回顾性分析[J]. 中华流行病学杂志, 2014, 35 (10): 1131- 1134.
	FU Weijie , HU Maohong , LIU Xiaoqing , et al. Retrospective analysis of human avian influenza A(H10N8) virus infection in Jiangxi province[J]. Chinese Journal of Epidemiology, 2014, 35 (10): 1131- 1134.
4	Hu MH , Li XD , Ni XS , et al. Coexistence of avian influenza virus H10 and H9 subtypes among chickens in live poultry markets during an outbreak of infection with a novel H10N8 virus in humans in Nanchang China[J]. Jpn J Infect Dis, 2015, 68 (5): 364- 369. doi: 10.7883/yoken.JJID.2014.377
5	冯道斌, 宋文涛, 倪贤生, 等. 南昌市2014~2017年活禽市场禽流感病毒监测[J]. 中华实验和临床病毒学杂志, 2020, 34 (3): 264- 269. doi: 10.3760/cma.j.cn112866-20191210-00198
	FENG Daobin , SONG Wentao , NI Xiansheng , et al. Surveillance on the avian influenza virus in live poultry markets in Nanchang city from 2014 to 2017[J]. Chinese J Exp Clin Virol, 2020, 34 (3): 264- 269. doi: 10.3760/cma.j.cn112866-20191210-00198
6	Scott AB , Toribio JA , Singh M , et al. Low pathogenic avian influenza exposure risk assessment in Australian commercial chicken farms[J]. Front Vet Sci, 2018, 5, 68. doi: 10.3389/fvets.2018.00068
7	Zou S , Zhang Y , Li X , et al. Molecular characterization and receptor binding specificity of H9N2 avian influenza viruses based on poultry-related environmental surveillance in China between 2013 and 2016[J]. Virology, 2019, 529, 135- 143. doi: 10.1016/j.virol.2019.01.002
8	Wu JY , Lau EH , Yuan J , et al. Transmission risk of avian influenza virus along poultry supply chains in Guangdong, China[J]. J Infect, 2019, 79 (1): 43- 48. doi: 10.1016/j.jinf.2019.05.006
9	Welkers MRA , Pawestri HA , Fonville JM , et al. Genetic diversity and host adaptation of avian H5N1 influenza viruses during human infection[J]. Emerg Microbes Infect, 2019, 8 (1): 262- 271. doi: 10.1080/22221751.2019.1575700
10	Zhou JJ , Fang DY , Fu J , et al. Infection and replication of avian influenza H5N1 virus in an infected human[J]. Virus Genes, 2009, 39 (1): 76- 80. doi: 10.1007/s11262-009-0365-y
11	Zhang Z , Li R , Jiang LF , et al. The complexity of human infected AIV H5N6 isolated from China[J]. BMC Infect Dis, 2016, 16 (1): 600. doi: 10.1186/s12879-016-1932-1
12	Wei SH , Yang JR , Wu HS , et al. Human infection with avian influenza A H6N1 virus: an epidemiological analysis[J]. Lancet Respir Med, 2013, 1 (10): 771- 778. doi: 10.1016/S2213-2600(13)70221-2
13	Yuan J , Zhang L , Kan XZ , et al. Origin and molecular characteristics of a novel 2013 avian influenza A(H6N1) virus causing human infection in Taiwan[J]. Clin Infect Dis, 2013, 57 (9): 1367- 1368. doi: 10.1093/cid/cit479
14	Wu XX , Xiao LL , Li LJ . Research progress on human infection with avian influenza H7N9[J]. Front Med, 2020, 14 (1): 8- 20. doi: 10.1007/s11684-020-0739-z
15	Wang W , Artois J , Wang XL , et al. Effectiveness of live poultry market interventions on human infection with avian influenza A(H7N9) virus, China[J]. Emerg Infect Dis, 2020, 26 (5): 891- 901. doi: 10.3201/eid2605.190390
16	李保娣, 何军, 李红育, 等. 甘肃省人感染和外环境来源的H9N2禽流感病毒基因特征分析[J]. 中华流行病学杂志, 2020, 41 (8): 1345- 1351. doi: 10.3760/cma.j.cn112338-20190712-00518
	LI Baodi , HE Jun , LI Hongyu , et al. Analysis on genetic characteristics of H9N2 avian influenza virus isolated from human infection and external environment in Gansu province[J]. Chinese Journal of Epidemiology, 2020, 41 (8): 1345- 1351. doi: 10.3760/cma.j.cn112338-20190712-00518
17	Xu JN , Li S , Yang Y , et al. Human infection with a further evolved avian H9N2 influenza A virus in Sichuan, China[J]. Sci China Life Sci, 2018, 61 (5): 604- 606. doi: 10.1007/s11427-017-9150-8
18	Zhang T , Bi YH , Tian HY , et al. Human infection with influenza virus A(H10N8) from live poultry markets, China, 2014[J]. Emerg Infect Dis, 2014, 20 (12): 2076- 2079.
19	Zhang W , Wan JG , Qian KJ , et al. Clinical characteristics of human infection with a novel avian-origin influenza A(H10N8) virus[J]. Chin Med J (Engl), 2014, 127 (18): 3238- 3242. doi: 10.1097/00029330-201409200-00009
20	Wen F , Li WF , Guo JY , et al. Genetic characterization of a novel genotype H9N2 avian influenza virus from chicken in South China[J]. J Infect, 2020, 81 (5): 816- 846.
21	Li X , Chen JL , Jin XJ , et al. Characterization of three H3N2 and one new reassortant H3N8 avian influenza virus in South China[J]. Infect Genet Evol, 2019, 75, 104016. doi: 10.1016/j.meegid.2019.104016
22	Hu JK , Zha YF , Jin XJ , et al. Continuous adaptation of the HA and NA gene of H3N2 subtypes of avian influenza virus in South China, 2017-2018[J]. J Infect, 2019, 79 (1): 61- 74.
23	Kang N , Chen M , Bi FY , et al. First positive detection of H9 Subtype of avian influenza virus nucleic acid in aerosol samples from live poultry markets in Guangxi, south of China[J]. Chin Med J (Engl), 2016, 129 (11): 1371- 1373. doi: 10.4103/0366-6999.182824
24	Fu XL , Huang YM , Fang B , et al. Evidence of H10N8 influenza virus infection among swine in southern China and its infectivity and transmissibility in swine[J]. Emerg Microbes Infect, 2020, 9 (1): 88- 94. doi: 10.1080/22221751.2019.1708811
25	Senne DA , Panigrahy B , Kawaoka Y , et al. Survey of the hemagglutinin (HA) cleavage site sequence of H5 and H7 avian influenza viruses: amino acid sequence at the HA cleavage site as a marker of pathogenicity potential[J]. Avian Dis, 1996, 40 (2): 425- 437. doi: 10.2307/1592241
26	Van Riel D , Munster VJ , De Wit E , et al. H5N1 virus attachment to lower respiratory tract[J]. Science, 2006, 312 (5772): 399. doi: 10.1126/science.1125548
27	Liu X , Yang C , Sun X , et al. Evidence for a novel mechanism of influenza A virus host adaptation modulated by PB2-627[J]. FEBS J, 2019, 286 (17): 3389- 3400. doi: 10.1111/febs.14867
28	McGuire KL , Smit P , Ess DH , et al. Mechanism and kinetics of copper complexes binding to the influenza A M2 S31N and S31N/G34E channels[J]. Biophys J, 2021, 120 (1): 168- 177. doi: 10.1016/j.bpj.2020.11.016
29	Tzitzoglaki C , McGuire K , Lagarias P , et al. Chemical probes for blocking of influenza A M2 wild-type and S31N channels[J]. ACS Chem Biol, 2020, 15 (9): 2331- 2337. doi: 10.1021/acschembio.0c00553
30	Cho JH , Zhao B , Shi J , et al. Molecular recognition of a host protein by NS1 of pandemic and seasonal influenza A viruses[J]. Proc Natl Acad Sci USA, 2020, 117 (12): 6550- 6558. doi: 10.1073/pnas.1920582117

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毒株名称	缩写	亚型	采样日期	采样场所	样本种类
A/chicken/Jiangxi/24472/2014	JX24472	H7/H10/N9	2014-04-29	活禽零售市场	鸡泄殖腔
A/environment/Jiangxi/07531/2015	JX07531	H10N3	2015-11-19	鸭养殖场	鸭粪便
A/environment/Jiangxi/07505/2015	JX07505	H10N3	2015-11-19	鸭养殖场	鸭笼具
A/environment /Jiangxi/07518/2015	JX07518	H10N3	2015-11-19	鸭养殖场	鸭粪便
A/environment /Jiangxi/07527/2015	JX07527	H10N3	2015-11-19	鸭养殖场	鸭粪便
A/environment/ Jiangxi/12590/2019	JX12590	H10N3	2019-06-18	批发市场	鸭粪便
A/environment/Guangxi/13425/2018	GX13425	H10N3	2018-12-24	活禽零售市场	脱毛机
A/environment/Guangxi/25019/2019	GX25019	H10N3	2019-01-02	活禽零售市场	鸭粪便
A/environment/Hunan/12257/2019	HN12257	H10N8	2019-08-19	活禽零售市场	禽咽拭子
A/environment/Hunan/12259/2019	HN12259	H10N8	2019-08-19	活禽零售市场	禽咽拭子
A/environment/Hunan/12263/2019	HN12263	H10N8	2019-08-19	活禽零售市场	鸭笼具

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