16株新型H3N3亚型禽流感病毒的遗传变异分析

doi:10.11843/j.issn.0366-6964.2024.09.027

摘要/Abstract

摘要：

为了解新出现的H3N3亚型禽流感病毒(AIV)的遗传变异特征, 对2022—2023年分离鉴定的H3亚型AIV进行了全基因组测序分析, 并对代表株进行了抗原差异分析。结果共分离鉴定出15株家禽源和1株野鸟源H3N3亚型AIV。全基因遗传演化分析结果显示, 16株分离株均属于欧亚分支, 其HA基因均源自H3N8亚型AIV, 与人源H3N8亚型AIV的核苷酸相似性为97.3%~99.2%;NA基因均源自H10N3亚型AIV, 与人源H10N3亚型毒株的核苷酸相似性为98.1%~98.4%;内部基因片段均来自H9N2亚型AIV。分离株HA基因裂解位点均符合低致病性AIV的分子特征。分离毒株存在多个能增强病毒对哺乳动物适应性及致病性的突变位点, 如PB2蛋白的A588V和E627V突变、PB1蛋白的I368V和S375N突变等。抗原差异性分析结果表明, 分离株与华东地区早期流行株的HI抗体滴度相差1 log2~3 log2。因此, 本研究分离的16株H3N3亚型AIV均为三源重组病毒, 由H9N2亚型AIV提供全部内部基因, 毒株存在多个哺乳动物适应性及致病性增强的突变, 并已传播给野禽。本研究揭示了新型H3N3亚型AIV的遗传特征、变异和多宿主分布情况, 为禽流感的防控提供理论支持。

关键词: 禽流感病毒, 新型, H3N3亚型, 基因变异, 抗原变异

Abstract:

In order to understand the genetic variation characteristics of the newly emerged H3N3 subtype avian influenza virus (AIV), we conducted whole-genome sequence analysis of H3 subtype AIV isolated during 2022-2023, and analyzed antigenic differences among representative strains. A total of 15 strains of H3N3 subtype AIV from poultry origin and 1 strain from wildfowl origin were isolated and identified. Phylogenetic analysis of the whole genome revealed that all 16 isolates belonged to the Eurasian lineage, with the HA gene derived from H3N8 subtype AIV, sharing a nucleotide homology of 97.3% to 99.2% when compared to human H3N8 subtype AIV isolates. The NA gene was derived from H10N3 subtype AIV, with a nucleotide homology of 98.1% to 98.4% when compared to human H10N3 subtype AIV isolates. The internal gene segments were all derived from H9N2 subtype AIVs. The cleavage sites of the HA gene of the isolates are consistent with the molecular characteristics of low pathogenic AIV. The isolated strains have multiple mutation sites that can enhance the adaptability and pathogenicity of the virus in mammals, such as the A588V and E627V mutations in the PB2 protein, the I368V and S375N mutations in the PB1 protein. Antigenic difference analysis showed that the isolates had HI antibody titers 1 log2-3 log2 different from those of earlier circulating strains in East China. Therefore, all 16 strains of H3N3 AIVs in this study are triple-recombinant viruses, with H9N2 AIV serving as the donor for all internal genes, and they harbor multiple amino acid mutations that enhance their adaptability and pathogenicity in mammals. These strains have already spread to wildfowl. This study reveals the genetic characteristics, variation and multi host distribution of the novel H3N3 AIVs, providing theoretical support for the prevention and control of avian influenza.

Key words: avian influenza virus, novel type, H3N3 subtype, genetic variation, antigenic variation

中图分类号:

S852.659.5

赵康宁, 杨忠龙, 陈怡, 朱春成, 郭云飞, 印云聪, 秦涛, 陈素娟, 彭大新. 16株新型H3N3亚型禽流感病毒的遗传变异分析[J]. 畜牧兽医学报, 2024, 55(9): 4029-4040.

Kangning ZHAO, Zhonglong YANG, Yi CHEN, Chuncheng ZHU, Yunfei GUO, Yuncong YIN, Tao QIN, Sujuan CHEN, Daxin PENG. Genetic Variation Analysis of Sixteen Novel H3N3 Subtype Avian Influenza Viruses[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 4029-4040.

图/表 4

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参考文献 57

1	周勇, 李知新, 鲁宏伟, 等. 我国H5和H7N9亚型高致病性禽流感的监测及疫情暴发分析[J]. 畜牧兽医学报, 2022, 53 (9): 3093- 3106. doi: 10.11843/j.issn.0366-6964.2022.09.024
	ZHOU Y , LI Z X , LU H W , et al. Surveillance and outbreak analysis of H5 and H7N9 subtypes of highly pathogenic avian influenza in China[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53 (9): 3093- 3106. doi: 10.11843/j.issn.0366-6964.2022.09.024
2	SPACKMAN E . A brief introduction to avian influenza virus[J]. Methods Mol Biol, 2020, 2123, 83- 92.
3	SPACKMAN E , KILLIAN M L . Avian influenza virus isolation, propagation, and titration in embryonated chicken eggs[J]. Methods Mol Biol, 2020, 2123, 149- 164.
4	OLSEN B , MUNSTER V J , WALLENSTEN A , et al. Global patterns of influenza a virus in wild birds[J]. Science, 2006, 312 (5772): 384- 388. doi: 10.1126/science.1122438
5	YANG J Y , ZHANG Y , YANG L , et al. Evolution of avian influenza virus (H3) with spillover into humans, China[J]. Emerg Infect Dis, 2023, 29 (6): 1191- 1201.
6	YANG J Y , YANG L , ZHU W F , et al. Epidemiological and genetic characteristics of the H3 subtype avian influenza viruses in China[J]. China CDC Wkly, 2021, 3 (44): 929- 936. doi: 10.46234/ccdcw2021.225
7	钱忠明. 华东地区家鸭中不同HA亚型禽流感病毒的分布以及从家养水禽和鸡中分离的H5亚型禽流感病毒HA基因的分子流行病学[D]. 扬州: 扬州大学, 2004.
	QIAN Z M. Distribution of avian influenza viruses of different HA subtypes among domestic ducks in eastern China and molecular epidemiology of HA genes of H5 subtype avian influenza viruses isolated from domestic waterfowls and chickens[D]. Yangzhou: Yangzhou University, 2004. (in Chinese)
8	QIU B F , LIU W J , PENG D X , et al. A reverse transcription-PCR for subtyping of the neuraminidase of avian influenza viruses[J]. J Virol Methods, 2009, 155 (2): 193- 198. doi: 10.1016/j.jviromet.2008.10.001
9	蒋梅. 禽流感病毒血凝素基因研究进展[J]. 中国人兽共患病学报, 2013, 29 (8): 817- 820. doi: 10.3969/cjz.j.issn.1002-2694.2013.08.017
	JIANG M . Research advances on hemagglutinin (HA) gene of avian influenza virus (AIV)[J]. Chinese Journal of Zoonoses, 2013, 29 (8): 817- 820. doi: 10.3969/cjz.j.issn.1002-2694.2013.08.017
10	BOSCH F X , GARTEN W , KLENK H D , et al. Proteolytic cleavage of influenza virus hemagglutinins: primary structure of the connecting peptide between HA1 and HA2 determines proteolytic cleavability and pathogenicity of Avian influenza viruses[J]. Virology, 1981, 113 (2): 725- 735. doi: 10.1016/0042-6822(81)90201-4
11	WILEY D C , WILSON I A , SKEHEL J J . Structural identification of the antibody-binding sites of Hong Kong influenza haemagglutinin and their involvement in antigenic variation[J]. Nature, 1981, 289 (5796): 373- 378. doi: 10.1038/289373a0
12	COLMAN P M , HOYNE P A , LAWRENCE M C . Sequence and structure alignment of paramyxovirus hemagglutinin-neuraminidase with influenza virus neuraminidase[J]. J Virol, 1993, 67 (6): 2972- 2980. doi: 10.1128/jvi.67.6.2972-2980.1993
13	COLMAN P M , VARGHESE J N , LAVER W G . Structure of the catalytic and antigenic sites in influenza virus neuraminidase[J]. Nature, 1983, 303 (5912): 41- 44. doi: 10.1038/303041a0
14	CHEN H L , BRIGHT R A , SUBBARAO K , et al. Polygenic virulence factors involved in pathogenesis of 1997 Hong Kong H5N1 influenza viruses in mice[J]. Virus Res, 2007, 128 (1-2): 159- 163. doi: 10.1016/j.virusres.2007.04.017
15	PATERSON D , TE VELTHUIS A J W , VREEDE F T , et al. Host restriction of influenza virus polymerase activity by PB2 627E is diminished on short viral templates in a nucleoprotein-independent manner[J]. J Virol, 2014, 88 (1): 339- 344. doi: 10.1128/JVI.02022-13
16	HSIA H P , YANG Y H , SZETO W C , et al. Amino acid substitutions affecting aspartic acid 605 and valine 606 decrease the interaction strength between the influenza virus RNA polymerase PB2 '627' domain and the viral nucleoprotein[J]. PLoS One, 2018, 13 (1): e0191226. doi: 10.1371/journal.pone.0191226
17	程善菊, 李金平, 侯广宇, 等. 14株H3亚型禽流感病毒全基因组序列分析[J]. 中国兽医杂志, 2018, 54 (10): 30- 35.
	CHENG S J , LI J P , HOU G Y , et al. Whole genome sequence analysis of 14 strains of H3 subtype avian influenza virus[J]. Chinese Journal of Veterinary Medicine, 2018, 54 (10): 30- 35.
18	刘开拓, 高如一, 王晓泉, 等. 新型重组H10N3亚型禽流感病毒对公共卫生安全的威胁[J]. 病毒学报, 2022, 38 (4): 958- 964.
	LIU K T , GAO R Y , WANG X Q , et al. Threat posed by a novel reassortant avian influenza virus (H10N3) to public health[J]. Chinese Journal of Virology, 2022, 38 (4): 958- 964.
19	裴瑞青, 张如胜, 叶文, 等. 湖南省首例人感染H3N8亚型禽流感病例的病毒分离及分子进化分析[J]. 中国人兽共患病学报, 2023, 39 (4): 364- 375.
	PEI R Q , ZHANG R H , YE W , et al. Viral isolation and molecular evolution analysis of the first human case of H3N8 subtype avian influenza infection in Hunan Province[J]. Chinese Journal of Zoonoses, 2023, 39 (4): 364- 375.
20	CHENG D L , DONG Y L , WEN S F , et al. A child with acute respiratory distress syndrome caused by avian influenza H3N8 virus[J]. J Infect, 2022, 85 (2): 174- 211.
21	张醒海. 候鸟禽流感病毒遗传变异规律分析及在哺乳动物间传播机制研究[D]. 吉林: 吉林大学, 2021.
	ZHANG X H. Genetic analysis and transmission mechanisms among mammals of migratory bird-origin avian influenza virus[D]. Jilin: Jilin University, 2021. (in Chinese)
22	何世成, 彭志, 王卫国, 等. 2011—2015年环洞庭湖区H3亚型禽流感病毒的分离鉴定与遗传演化[J]. 畜牧兽医学报, 2019, 50 (2): 382- 389. doi: 10.11843/j.issn.0366-6964.2019.02.016
	HE S C , PENG Z , WANG W G , et al. Isolation, identification and genetic evolution of H3 subtype avian influenza virus in dongting lake region from 2011 to 2015[J]. Acta Veterinaria et Zootechnica Sinica, 2019, 50 (2): 382- 389. doi: 10.11843/j.issn.0366-6964.2019.02.016
23	GAO R Y , ZHENG H F , LIU K T , et al. Genesis, evolution and host species distribution of influenza A (H10N3) virus in China[J]. J Infect, 2021, 83 (5): 607- 635.
24	GU M , CHEN H Z , LI Q H , et al. Enzootic genotype S of H9N2 avian influenza viruses donates internal genes to emerging zoonotic influenza viruses in China[J]. Vet Microbiol, 2014, 174 (3-4): 309- 315. doi: 10.1016/j.vetmic.2014.09.029
25	申松玮, 王泽源, 范威峰, 等. 2017—2018年华东地区H9亚型禽流感病毒分离毒株的抗原差异分析[J]. 畜牧兽医学报, 2019, 50 (6): 1230- 1238. doi: 10.11843/j.issn.0366-6964.2019.06.013
	SHEN S W , WANG Z Y , FAN W F , et al. Antigen difference analysis of H9 subtype avian influenza viruses isolated in east China during 2017-2018[J]. Acta Veterinaria et Zootechnica Sinica, 2019, 50 (6): 1230- 1238. doi: 10.11843/j.issn.0366-6964.2019.06.013
26	ZHAO G , GU X B , LU X L , et al. Novel reassortant highly pathogenic H5N2 avian influenza viruses in poultry in China[J]. PLoS One, 2012, 7 (9): e46183. doi: 10.1371/journal.pone.0046183
27	ZHANG Q Y , SHI J Z , DENG G H , et al. H7N9 influenza viruses are transmissible in ferrets by respiratory droplet[J]. Science, 2013, 341 (6144): 410- 414. doi: 10.1126/science.1240532
28	许冠龙, 张谞霄, 孙洪磊, 等. H10N7亚型禽流感病毒鼠适应毒的致病机制研究[C]//第四届全国禽病分子生物技术青年工作者会议. 天津: 中国畜牧兽医学会禽病学分会, 2015.
	XU G L, ZHANG X X, SUN H L, et al. Substitutions in PB2 and NA contributes to the pathogenicity and neurovirulence of H10N7 influenza virus[C]//Proceedings of the 4th National Conference of Young Scientists in Avian Disease Molecular Biotechnology. Tianjin, 2015. (in Chinese)
29	LI J W , ISHAQ M , PRUDENCE M , et al. Single mutation at the amino acid position 627 of PB2 that leads to increased virulence of an H5N1 avian influenza virus during adaptation in mice can be compensated by multiple mutations at other sites of PB2[J]. Virus Res, 2009, 144 (1-2): 123- 129. doi: 10.1016/j.virusres.2009.04.008
30	崔鹏飞. 2009—2013年中国H3N8亚型禽流感病毒生物学特性研究[D]. 兰州: 甘肃农业大学, 2016.
	CUI P F. Biological characterization of H3N8 avian influenza viruses isolated from 2009 to 2013 in China[D]. Lanzhou: Gansu Agricultural University, 2016. (in Chinese)
31	CONENELLO G M , ZAMARIN D , PERRONE L A , et al. A single mutation in the PB1-F2 of H5N1 (HK/97) and 1918 influenza A viruses contributes to increased virulence[J]. PLoS Pathog, 2007, 3 (10): 1414- 1421.
32	HU M , CHU H , ZHANG K , et al. Amino acid substitutions V63I or A37S/I61T/V63I/V100A in the PA N-terminal domain increase the virulence of H7N7 influenza A virus[J]. Sci Rep, 2016, 6 (1): 37800. doi: 10.1038/srep37800
33	LIANG L B , JIANG L , LI J P , et al. Low polymerase activity attributed to PA drives the acquisition of the PB2 E627K mutation of H7N9 avian influenza virus in mammals[J]. mBio, 2019, 10 (3): e01162- 19.
34	SONG W J , WANG P , MOK B W Y , et al. The K526R substitution in viral protein PB2 enhances the effects of E627K on influenza virus replication[J]. Nat Commun, 2014, 5 (1): 5509. doi: 10.1038/ncomms6509
35	LUK G S M , LEUNG C Y H , SIA S F , et al. Transmission of H7N9 influenza viruses with a polymorphism at PB2 residue 627 in chickens and ferrets[J]. J Virol, 2015, 89 (19): 9939- 9951. doi: 10.1128/JVI.01444-15
36	LI O T W , CHAN M C W , LEUNG C S W , et al. Full factorial analysis of mammalian and avian influenza polymerase subunits suggests a role of an efficient polymerase for virus adaptation[J]. PLoS One, 2009, 4 (5): e5658. doi: 10.1371/journal.pone.0005658
37	PENA L , VINCENT A L , LOVING C L , et al. Restored PB1-F2 in the 2009 pandemic H1N1 influenza virus has minimal effects in swine[J]. J Virol, 2012, 86 (10): 5523- 5532. doi: 10.1128/JVI.00134-12
38	DURAIRAJ K , TRINH T T T , YUN S Y , et al. Molecular characterization and pathogenesis of h6n6 low pathogenic avian influenza viruses isolated from mallard ducks (Anas platyrhynchos) in South Korea[J]. Viruses, 2022, 14 (5): 1001. doi: 10.3390/v14051001
39	CHEN H Y , YUAN H , GAO R B , 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
40	HULSE-POST D J , FRANKS J , BOYD K , et al. Molecular changes in the polymerase genes (PA and PB1) associated with high pathogenicity of H5N1 influenza virus in mallard ducks[J]. J Virol, 2007, 81 (16): 8515- 8524. doi: 10.1128/JVI.00435-07
41	YAMAYOSHI S , YAMADA S , FUKUYAMA S , et al. Virulence-affecting amino acid changes in the PA protein of H7N9 influenza A viruses[J]. J Virol, 2014, 88 (6): 3127- 3134. doi: 10.1128/JVI.03155-13
42	DE WIT E , MUNSTER V J , VAN RIEL D , et al. Molecular determinants of adaptation of highly pathogenic avian influenza H7N7 viruses to efficient replication in the human host[J]. J Virol, 2010, 84 (3): 1597- 1606. doi: 10.1128/JVI.01783-09
43	DUCATEZ M F , ILYUSHINA N A , FABRIZIO T P , et al. Both influenza hemagglutinin and polymerase acidic genes are important for delayed pandemic 2009 H1N1 virus clearance in the ferret model[J]. Virology, 2012, 432 (2): 389- 393. doi: 10.1016/j.virol.2012.06.018
44	SONG J S , FENG H P , XU J , et al. The PA protein directly contributes to the virulence of H5N1 avian influenza viruses in domestic ducks[J]. J Virol, 2011, 85 (5): 2180- 2188. doi: 10.1128/JVI.01975-10
45	ZHENG M , LUO J , CHEN Z . Development of universal influenza vaccines based on influenza virus M and NP genes[J]. Infection, 2014, 42 (2): 251- 262. doi: 10.1007/s15010-013-0546-4
46	WASILENKO J L , LEE C W , SARMENTO L , et al. NP, PB1, and PB2 viral genes contribute to altered replication of H5N1 avian influenza viruses in chickens[J]. J Virol, 2008, 82 (9): 4544- 4553. doi: 10.1128/JVI.02642-07
47	TADA T , SUZUKI K , SAKURAI Y , et al. NP body domain and PB2 contribute to increased virulence of H5N1 highly pathogenic avian influenza viruses in chickens[J]. J Virol, 2011, 85 (4): 1834- 1846. doi: 10.1128/JVI.01648-10
48	ILYUSHINA N A , KHALENKOV A M , SEILER J P , et al. Adaptation of pandemic H1N1 influenza viruses in mice[J]. J Virol, 2010, 84 (17): 8607- 8616. doi: 10.1128/JVI.00159-10
49	SHI J Z , DENG G H , KONG H H , et al. H7N9 virulent mutants detected in chickens in China pose an increased threat to humans[J]. Cell Res, 2017, 27 (12): 1409- 1421. doi: 10.1038/cr.2017.129
50	KAPLAN B S , KIMBLE J B , CHANG J , et al. Aerosol transmission from infected swine to ferrets of an H3N2 virus collected from an agricultural fair and associated with human variant infections[J]. J Virol, 2020, 94 (16): e01009- 20.
51	FAN S F , DENG G H , SONG J S , et al. Two amino acid residues in the matrix protein M1 contribute to the virulence difference of H5N1 avian influenza viruses in mice[J]. Virology, 2009, 384 (1): 28- 32. doi: 10.1016/j.virol.2008.11.044
52	SEO S H , HOFFMANN E , WEBSTER R G . RETRACTED: the NS1 gene of H5N1 influenza viruses circumvents the host anti-viral cytokine responses[J]. Virus Res, 2004, 103 (1-2): 107- 113. doi: 10.1016/j.virusres.2004.02.022
53	CHEN S J , MIAO X Y , HUANGFU D D , et al. H5N1 avian influenza virus without 80-84 amino acid deletion at the NS1 protein hijacks the innate immune system of dendritic cells for an enhanced mammalian pathogenicity[J]. Transbound Emerg Dis, 2021, 68 (4): 2401- 2413. doi: 10.1111/tbed.13904
54	MIN J Y , LI S D , SEN G C , et al. A site on the influenza A virus NS1 protein mediates both inhibition of PKR activation and temporal regulation of viral RNA synthesis[J]. Virology, 2007, 363 (1): 236- 243. doi: 10.1016/j.virol.2007.01.038
55	DRAKOPOULOS A , TZITZOGLAKI C , MCGUIRE K , et al. Unraveling the binding, proton blockage, and inhibition of influenza M2 WT and S31N by rimantadine variants[J]. ACS Med Chem Lett, 2018, 9 (3): 198- 203. doi: 10.1021/acsmedchemlett.7b00458
56	RAMSAY L C , BUCHAN S A , STIRLING R G , et al. Retraction note: the impact of repeated vaccination on influenza vaccine effectiveness: a systematic review and meta-analysis[J]. BMC Medicine, 2018, 16 (1): 133. doi: 10.1186/s12916-018-1137-0
57	SHIH A C C , HSIAO T C , HO M S , et al. Simultaneous amino acid substitutions at antigenic sites drive influenza A hemagglutinin evolution[J]. Proc Natl Acad Sci U S A, 2007, 104 (15): 6283- 6288. doi: 10.1073/pnas.0701396104

毒株名称	缩写	宿主	日期	来源
Virus names	Abbreviation	Host	Date	Source
A/China/ZMD-22-2/2022(H3N8)^*	ZMD-22-2	Human	2022/04	Human
A/China/CSKFQ-22-5/2022(H3N8)^*	CSKFQ-22-5	Human	2022/05	Human
A/China/0428/2021(H10N3)^*	0428	Human	2021/04	Human
A/duck/Fujian/SD063/2017(H3N3)^*	SD063	Duck	2017/03	Unknown
A/chicken/Jiangsu/W23910/2017(H3N2)^*	W23910	Chicken	2017/02	LBM
A/chicken/YangZhou/YZ12145/2022(H3N3)	YZ12145	Chicken	2022/12	LBM
A/chicken/YangZhou/YZ01082/2023(H3N3)	YZ01082	Chicken	2023/01	Specimen
A/chicken/NanTong/NT02101/2023(H3N3)	NT02101	Chicken	2023/02	Specimen
A/chicken/YangZhou/YZ02283/2023(H3N3)	YZ02283	Chicken	2023/02	LBM
A/chicken/YangZhou/YZ02284/2023(H3N3)	YZ02284	Chicken	2023/02	LBM
A/chicken/YangZhou/YZ02285/2023(H3N3)	YZ02285	Chicken	2023/02	LBM
A/chicken/YangZhou/YZ02288/2023(H3N3)	YZ02288	Chicken	2023/02	LBM
A/chicken/NanTong/NT030601/2023(H3N3)	NT030601	Chicken	2023/03	Specimen
A/duck/YangZhou/YZD031512/2023(H3N3)	YZ031512	Duck	2023/03	LBM
A/chicken/YangZhou/YZ031524/2023(H3N3)	YZ031524	Chicken	2023/03	LBM
A/chicken/YangZhou/YZ031542/2023(H3N3)	YZ031542	Chicken	2023/03	LBM
A/chicken/YangZhou/YZ031554/2023(H3N3)	YZ031554	Chicken	2023/03	LBM
A/chicken/YangZhou/YZ032702/2023(H3N3)	YZ032702	Chicken	2023/03	Specimen
A/chicken/YangZhou/YZ042881/2023(H3N3)	YZ042881	Chicken	2023/04	LBM
A/chicken/YangZhou/YZ042893/2023(H3N3)	YZ042893	Chicken	2023/04	LBM
A/wildfowl/HuaDong/SY01/2023(H3N3)	SY01	Wildfowl	2023/01	Wildfowl

血清 Sera	病毒Virus
血清 Sera	A/chicken/Jiangsu/W23910/2017 (H3N2)	A/chicken/NanTong/NT02101/2023 (H3N3)	A/chicken/YangZhou/YZ01082/2023 (H3N3)	A/chicken/YangZhou/YZ032702/2023 (H3N3)
A/chicken/Jiangsu/W23910/2017(H3N2)^a	9.33±0.58^b	8.67±0.58	7.00±0.00	7.33±0.58
A/chicken/NanTong/NT02101/2023(H3N3)^a	6.33±0.58	9.00±0.00	7.75±0.50	8.00±0.00

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[11]	李静云, 连朋敬, 白玉, 奚柳青, 张子卉, 牛小飞, 杨俊琦, 乔健. H9N2亚型禽流感病毒感染对小鼠肠道菌群的影响[J]. 畜牧兽医学报, 2021, 52(5): 1359-1368.
[12]	李丽, 唐国毅, 冯贺龙, 薛玉涵, 任助, 王国康, 贾妙妙, 商雨, 罗青平, 邵华斌, 温国元. 基于马赛克HA序列的H9亚型禽流感灭活疫苗的免疫效力分析[J]. 畜牧兽医学报, 2021, 52(12): 3569-3577.
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[14]	戴学宇, 张乾义, 徐璐, 赵启祖, 王琴, 夏应菊. CRISPR/Cas9基因编辑技术在重要猪病毒病防控中的研究与应用[J]. 畜牧兽医学报, 2020, 51(5): 943-951.
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