1株欧亚类禽猪流感病毒的分离鉴定及遗传进化分析

doi:10.16431/j.cnki.1671-7236.2024.04.032

Abstract

Abstract: 【Objective】 The experiment was aimed to understand the prevalence of Swine influenza virus (SIV) in Guangdong and explore its molecular biological characteristics.【Method】 Nasal swab and lung tissue samples of pigs suspected to be infected with Swine influenza virus were collected from a pig farm in Guangdong for virus isolation and identification,genetic evolution and key amino acid site analysis.【Result】 The samples were tested positive for Swine influenza virus nucleic acid by Real-time quantitative RT-PCR.In the red blood cell agglutination test,the virus exhibited agglutination with chicken red blood cells,with a hemagglutination titer of 1∶128.The results of sequencing analysis of eight gene segments were analyzed by BLAST comparison and construction of evolutionary trees,and HA and NA genes belonged to the Eurasian avian-like Swine influenza virus (H1N1) branch,PA,PB1,PB2,NP and M genes belonged to the pdm/09 branch,and NS gene belonged to the North American triple reassortant branch,therefore,the isolate in this study belonged to the G4 genotype Eurasian avian-like Swine influenza virus,and was named A/swine/Guangdong/CJM2/2022 (H1N1).Key amino acid site analysis showed that the cleavage site sequence of HA protein of the isolate was PSIQSR/GL,which had the molecular characteristics of a typical low pathogenicity influenza virus.The amino acids at position 190,225 and 226 of the receptor binding site of HA gene were D,E and Q,respectively,which suggested that it had the potential ability to bind to the human salivary acid receptor and it also had the potential to bind avian sialic acid receptors.None of the amino acid residues of NA gene were mutated,suggesting that the isolate was sensitive to neuraminidase inhibitors such as oseltamivir and zanamivir.However,the mutations at three amino acid sites in M gene were V27A,A30T and D44A,suggesting an increased resistance to amantadine-based drugs.【Conclusion】 The strain isolated and identified in this study was a G4 Eurasian avian-like Swine influenza virus.The analysis of key amino acid sites suggested that this strain had the characteristics of adapting to replication and enhancing virulence in mammals.The results of this study provided reference data for the prevention and control of swine influenza in Guangdong.

Key words: Eurasian avian-like Swine influenza virus; isolation and identification; sequence analysis; H1N1 subtype

CLC Number:

S852.65⁺1

HAN Hui, GUO Yajing, YAN Guangzhi, CHEN Shengnan, LIU Mingjie, MO Meilian, HUANG Liangzong. Isolation,Identification and Genetic Evolution Analysis of a Eurasian Avian-like Swine Influenza Virus[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1642-1650.

References

[1] 黄良宗,颜广智,邓汝森,等.猪流感病毒广东株分离鉴定及遗传进化分析[J].中国畜牧兽医,2020,47(8):2625-2633. HUANG L Z,YAN G Z,DENG R S,et al.Isolation,identification and genetic evolution analysis of Swine influenza virus from Guangdong province[J].China Animal Husbandry & Veterinary Medicine,2020,47(8):2625-2633.(in Chinese)
[2] ITO T,COUCEIRO J N,KElM S,et al.Molecular basis for the generation in pigs of Influenza A viruses with pandemic potential[J]. Journal of Virology,1998,72(9):7367-7373.
[3] MA W,KAHN R E,RICHT J A.The pig as a mixing vessel for Influenza viruses:Human and veterinary implications[J].Journal of Molecular and Genetic Medicine,2008,3(1):158-166.
[4] PENSAERT M,OTTIS K,VANDEPUTT J,et al.Evidence for the natural transmission of Influenza A virus from wild ducts to swine and its potential importance for man[J].Bulletin of the World Health Organization,1981,59(1):75-78.
[5] LI H,LENG H,TANG S,et al.Prevalence,genetics and evolutionary properties of Eurasian avian-like H1N1 Swine influenza viruses in Liaoning[J].Viruses,2022,14(3):643.
[6] CENTERS FOR DISEASE CONTROL AND PREVENTION (CDC).Update:Swine influenza A (H1N1) infections—California and Texas,April 2009[J]. Morbidity and Mortality Weekly Report,2009,58(16):435-437.
[7] VIJAYKRISHNA D,POON LL,ZHU H C,et al.Reassortment of pandemic H1N1/2009 Influenza A virus in swine[J].Science,2010,328(5985):1529.
[8] SUN H,XIAO Y,LIU J,et al.Prevalent Eurasian avian-like H1N1 Swine influenza virus with 2009 pandemic viral genes facilitating human infection[J]. Proceedings of the National Academy of Sciences of the United States of America,2022,117(29):17204-17210.
[9] 李梓,赵晓南,黄维娟,等.云南省首例人感染G4基因型欧亚类禽H1N1猪流感病毒病原学特征分析[J].病毒学报,2022,38(2):290-297. LI Z,ZHAO X N,HUANG W J,et al.Etiological characteristics of the first human infection with the G4 genotype Eurasian avian-like H1N1 Swine influenza virus in Yunnan province,China[J].Chinese Journal of Virology,2022,38(2):290-297.(in Chinese)
[10] MENG F,CHEN Y,SONG Z,et al.Continued evolution of the Eurasian avian-like H1N1 Swine influenza viruses in China[J].Science China Life Science, 2023,66(2):269-282.
[11] YANG H,CHEN Y,QIAO C,et al.Prevalence,genetics,and transmissibility in ferrets of Eurasian avian-like H1N1 Swine influenza viruses[J].Proceedings of the National Academy of Sciences of the United States of America,2016,113(2):392-397.
[12] 黄书,曾智勇,梁海英,等.1株H1N1亚型猪流感病毒的进化分析及分子特征研究[J].中国畜牧兽医,2023,50(12):5084-5093. HUANG S,ZENG Z Y,LIANG H Y,et al.Evolutionary analysis and molecular characteristics of a H1N1 subtype Swine influenza virus[J].China Animal Husbandry & Veterinary Medicine,2023,50(12):5084-5093.(in Chinese)
[13] 张春明,乔传玲,陈艳,等.猪流感病毒M基因实时荧光定量PCR诊断方法的建立[J].中国预防兽医学报,2008,30(10):805-810. ZHANG C M,QIAO C L,CHEN Y,et al.Development of Real-time fluorescent quantitative PCR for detection of M gene of Swine influenza viruses[J].Chinese Journal of Preventive Veterinary Medicine, 2008,30(10):805-810.(in Chinese)
[14] HOFFMANN E,STECH J,GUAN Y,et al.Universal primer set for the full-length amplification of all Influenza A viruses[J].Archives of Virology,2001,146(12):2275-2289.
[15] MCKIMM-BRESCHKIN J,TRIVEDI T,HAMPSON A,et al.Neuraminidase sequence analysis and susceptibilities of Influenza virus clinical isolates to zanamivir and oseltamivir[J].Antimicrobial Agents and Chemotherapy,2003,47(7):2264-2272.
[16] ZHU W,ZHANG H,XIANG X,et al.Reassortant Eurasian avian-like Influenza A(H1N1) virus from a severely ill child,Hunan province,China,2015[J].Emerging Infectious Diseases, 2016,22(11):1930-1936.
[17] QI X,CUI L,JIAO Y,et al.Antigenic and genetic characterization of a European avian-like H1N1 Swine influenza virus from a boy in China in 2011[J]. Archives of Virology,2013,158(1):39-53.
[18] WANG Z,YANG H,CHEN Y,et al.A single-amino-acid substitution at position 225 in hemagglutinin alters the transmissibility of avian-like H1N1 Swine influenza virus in guinea pigs[J].Journal of Virology,2017,91(21):e00800-17.
[19] GABRIEL G,DAUBER B,WOLFF T,et al.The viral polymerase mediates adaptation of an Avian influenza virus to a mammalian host[J].Proceedings of the National Academy of Sciences of the United States of America,2005,102(51):18590-18595.
[20] BUSSEY K A,BOUSSE T L,DESMET E A,et al.PB2 residue 271 plays a key role in enhanced polymerase activity of Influenza A viruses in mammalian host cells[J].Journal of Virology,2010,84(9):4395-4406.
[21] XU C,XU B,WU Y,et al.A single amino acid at position 431 of the PB2 protein determines the virulence of H1N1 Swine influenza viruses in mice[J].Journal of Virology,2020,94(8):e01930-19.
[22] SUN Y,XU Q,SHEN Y,et al.Naturally occurring mutations in the PA gene are key contributors to increased virulence of pandemic H1N1/09 Influenza virus in mice[J].Journal of Virology,2014,88(8):4600-4604.
[23] ZHU W,FENG Z,CHEN Y,et al.Mammalian-adaptive mutation NP-Q357K in Eurasian H1N1 Swine influenza viruses determines the virulence phenotype in mice[J]. Emerging Microbes & Infections,2019,8(1):989-999.
[24] FURUSE Y,SUZUKI A,OSHIANI H.Large-scale sequence analysis of M gene of Influenza A viruses from different species:Mechanisms for emergence and spread of amantadine resistance[J]. Antimicrobial Agents and Chemotherapy,2009,53(10):4457-4463.
[25] ZHU H,ZHOU B,FAN X,et al.Novel reassortment of Eurasian avian-like and pandemic/2009 Influenza viruses in swine:Infectious potential for humans[J]. Journal of Virology,2011,85(20):10432-10439.
[26] 郑运,马俊,罗永峰,等.广东省猪流感病毒的分离鉴定及遗传进化分析[J].中国兽医学报,2017,37(2):266-271. ZHENG Y,MA J,LUO Y F,et al.Isolation and genetic evolution analysis of Swine influenza virus in Guangdong province[J].Chinese Journal of Veterinary Science,2017,37(2):266-271.(in Chinese)

Isolation,Identification and Genetic Evolution Analysis of a Eurasian Avian-like Swine Influenza Virus

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