H9亚型禽流感病毒HF株的分离鉴定和免疫原性评价

doi:10.16431/j.cnki.1671-7236.2020.09.029

Abstract

Abstract: In 2015,an H9N2 subtype avian influenza virus (AIV) strain was isolated from a chicken farm in Hefei,Anhui,and named HF strain.The results of the chicken embryo proliferation characteristics study showed that the half infection rate of chicken embryo (EID₅₀) was 10^9.17/0.1 mL,and the mean time to death for minimum lethal dose(MDT) was 87 h.The analysis result of HA gene showed that its amino acid cleavage site was located in RSSR↓GLF,which accorded with the characteristics of low pathogenic avian influenza.The genetic evolution analysis of HA gene revealed that the isolate belonged to the h9.4.2.5 lineage,which accorded with the current virus strain epidemic characteristics.The HF strain was prepared with 10 H9N2 subtype AIV isolates which isolated from all over the country from 2006 to 2018 to prepare inactivated vaccines,immunize SPF chickens,prepare positive sera,and analyze the virus antigenicity by cross hemagglutination inhibition test.The results showed that the correlation between the HF strain and the virus antigens before 2014 and was between 0.50-0.56,and the virus antigen correlation after 2014 was 0.89-1.00.This showed that the isolate had good antigenic correlation with epidemic strains in recent years.Inactivate HF strain virus solution with 0.2% formaldehydel,and its HA titer did not change before and after inactivation.After the inactivated virus solution was prepared into an oil emulsion inactivated vaccine to immunize SPF chickens,21 days after immunization,the average value of the HI antibody titer reached 9.0log2.It could make immune chicken completely resistant to H9 subtype AIV infection and provide 100% protection from challenge.The above research results showed that the HF strain had good immunogenicity and could be used as a vaccine candidate strain for the prevention of H9N2 subtype AIV.

Key words: H9N2 subtype avian influenza virus; isolation and identification; genetic evolution; immunogenicity

CLC Number:

S852.65

CHENG Yi, TIAN Hui, ZHANG Ya, XUE Jingjing, SHENG Dongbei, LIU Wujie, WANG Wanbing, ZHANG Pantao, TIAN Kegong. Isolation,Identification and Immunogenicity Evaluation of H9N2 Subtype Avian Influenza Virus HF Strain[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2945-2952.

References

[1] 卡尔尼克B W.禽病学[M].第9版.高福,刘文军主译.北京:北京农业大学出版社,1991. CARNICK B W.Ornithology[M].9th Edition.GAO F,LIU W J (Translator).Beijing:Beijing Agricultural University Press,1991.(in Chinese)
[2] MUNSTER V J,FOUCHIER R A M.Avian influenza virus:Of virus and bird ecology[J].Vaccine,2009,27(45):6340-6344.
[3] CHEN H,SUBBARAO K,SWAYNE D,et al.Generation and evaluation of a high-growth reassortant H9N2 influenza A virus as a pandemic vaccine candidate[J].Vaccine,2003,21(17-18):1974-1979.
[4] 陈伯伦,张泽纪,陈伟斌.鸡A型禽流感病毒的分离与血清学初步鉴定[J].中国家禽,1997,11:5-7. CHEN B L,ZHANG Z J.Isolation and preliminary serological identification of avian influenza virus-type from chickens[J].China Poultry,1997,11:5-7.(in Chinese)
[5] 丛彦龙,钟颖,孙艺学,等.H9N2亚型禽流感病毒流行病学及其疫苗的研究进展[J].中国兽医学报,2017,37(2):386-392. CONG Y L,ZHONG Y,SUN Y X.Advance on epidemiology and vaccine of H9N2 avian influenza virus[J].Chinese Journal of Veterinary Science,2017,37(2):386-392.(in Chinese)
[6] WATANABE T,KISO M,FUKUYAMA S.et al.Characterization of H7N9 influenza A viruses isolated from humans[J].Nature,501(7468):551-555.
[7] SUN X,XU X,LIU Q,et al.Evidence of avian-like H9N2 influenza A virus among dogs in Guangxi,China[J].Infection Genetics & Evolution,2013,20(12):471-475.
[8] 钟功勋,李雁冰,平继辉,等.H9N2亚型禽流感病毒血凝素和神经氨酸酶基因的遗传分析[J].中国预防兽医学报,2007,29(12):946-949. ZHONG G X,LI Y B,PING J H,et al.Genetic analysis of the hemagglutinin(HA) and neuramini-dase(NA) genes of H9N2 influenza virus isolated from mainland China[J].Chinese Journal of Preventive Veterinary Medicine,2007,29(12):946-949.(in Chinese)
[9] 王泽霖,刘岩,李建丽,等.H9N2亚型禽流感病毒抗原性变异的研究[J].中国兽医学报,2007,27(3):351-354. WANG Z L,LIU Y,LI J L,et al.Antigenic drift of H9N2 subtype avian influenza virus strains isolated from different years[J].Chinese Journal of Veterinary Science,2007,27(3):351-354.(in Chinese)
[10] 孟芳,徐怀英,张伟,等.近20年中国部分地区鸡源H9N2亚型禽流感病毒HA基因遗传演化及其变异频率[J].微生物学报,2016,56(1):35-43. MENG F,XU H Y,ZHANG W,et al.Genetic evolution and substitution frequency of avian influenza virus HA gene in chicken H9N2 subtype in China in the last 20 years[J].Acta Microbiologica Sinica,2016,56(1):35-43.(in Chinese)
[11] 农业部兽用生物制品规程委员会.中华人民共和国兽用生物制品规程[M].北京:化学工业出版社,2000. VETERINARY BIOLOGICAL PRODUCTS REGULA-TION COMMITTEE OF THE MINISTRY OF AGRICULTURE.Regulations on Veterinary Biological Products of the People's Republic of China[M].Beijing:Chemical Industry Press,2000.(in Chinese)
[12] 杨汉春.动物免疫学[M].北京:中国农业大学出版社,1995. YANG H C.Animal Immunology[M].Beijing:China Agricultural University Press,1995.(in Chinese)
[13] 中国兽药典委员会.中华人民共和国兽药典[M].2015版.北京:中国农业出版社,2016. CHINESE VETERINARY PHARMACOPOEIA COMMITTEE.Veterinary Pharmacopoeia of the People's Republic of China[M].2015 Edition.Beijing:China Agricultural Press,2016.(in Chinese)
[14] SHUO L,KANG J,JIMING C,et al.Panorama phylogenetic diversity and distribution of type A influenza virus[J].PLoS One,2009,4(3):e5022.
[15] 葛菲菲,刘健,杨德全,等.H9N2亚型禽流感病毒抗原性变异的研究[J].中国动物传染病学报,2015,23(5):10-14. GE F F,LIU J,YANG D Q,et al.Antigenic drift of H9N2 subtype avian influenza virus strains isolated from different years[J].Chinese Journal of Animal Infectious Diseases,2015,23(5):10-14.(in Chinese)
[16] 张义冉,王贞,孙怡朋,等.2017年河北省蛋鸡养殖场H9N2亚型禽流感病毒的遗传演化和抗原性分析[J].微生物学报,2019,46(7):1803-1811. ZHANG Y R,WANG Z H,SUN Y P,et al.Genetic evolution and antigenic characterisitics of H9N2 avian influenza viruses in laying-hen farms in Hebei province in 2017[J].Microbiology China,2019,46(7):1803-1811.(in Chinese)
[17] 高斌,张帅,魏志鹏,等.2017~2018年山东省H9N2亚型禽流感病毒HA基因的遗传进化分析[J].中国家禽,2019,41(6):21-26. GAO B,ZHANG S H,WEI Z P,et al.Genetic evolution analysis of HA gene of H9N2 subtype avian influenza virus in Shandong province from 2017 to 2018[J].China Poultry,2019,41(6):21-26.(in Chinese)
[18] ZHU Y,YANG D,REN Q,et al.Identification and characterization of a novel antigenic epitope in the hemagglutinin of the escape mutants of H9N2 avian influenza viruses[J].Veterinary Microbiology,2015,178(1-2):144-149.
[19] 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. 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[J].Acta Veterinaria et Zootechnica Sinica,2019,50(6):1230-1238.(in Chinese)
[20] 尚飞雪,刘朔,蒋文明,等.近年来中国H9亚型禽流感分离株谱系分析[J].中国动物检疫,2012,29(4):51-53. SHANG F X,LIU S,JIANG W M.Phylogenetic analysis of H9 subtype avian influenza virus isolated from China in recent years[J].China Animal Health Inspection,2012,29(4):51-53.(in Chinese)
[21] HENRY K,CATHARINE W,HELENA L.et al.Genetic characterization and pathogenesis of the first H9N2 low pathogenic avian influenza viruses isolated from chickens in Kenyan live bird markets[J].Infection,Genetics and Evolution,2020,78:104074.
[22] JIANG W M,LIU S,KOU G Y,et al.Chinese and global distribution of H9 subtype avian influenza viruses[J].PLoS One,2012,7(12):e52671.
[23] 任助,商雨,薛玉涵,等.湖北地区鸡源H9N2亚型禽流感病毒的分离鉴定及遗传进化分析[J].动物医学进展,2019,40(6):30-36. REN Z,SHANG Y,XUE Y H,et al.Isolation,identification and genetic evolution analysis of chicken H9N2 avian influenza virus in Hubei province[J].Progress Veterinary Medicine,2019,40(6):30-36.(in Chinese)
[24] 陈春丽,王强,李群芳,等.一株H9亚型禽流感病毒的分离鉴定及生物特性研究[J].中国畜牧兽医,2015,42(10):2619-2624. CHEN C L,WANG Q,LI Q F,et al.Isolation and identification of one H9 subtype avian influenza virus and study on its biological characteristics[J].China Animal Husbandry & Veterinary Medicine,2015,42(10):2619-2624.(in Chinese)
[25] 杨志远,林健,王小蕾,等.一株H9N2亚型禽流感病毒的鉴定与HA基因序列分析[J].动物医学进展,2019,40(9):13-18. YANG Z Y,LIN J,WANG X L,et al.Identification and HA gene sequence analysis of a H9N2 subtype avian influenza virus isolate[J].Progress Veterinary Medicine,2019,40(9):13-18.(in Chinese)
[26] 卞学兵,曾嘉梅,周继勇,等.免疫鸡群中分离的H9N2亚型禽流感病毒的分子特征研究[J].中国预防兽医学报,2012,34:15-18. BIAN X B,ZENG J M,ZHOU J Y,et al.Molecular characterization of a H9N2 subtype avian influenza virus from the vaccinated chicken flock[J].Chinese Journal of Preventive Veterinary Medicine,2012,34:15-18.(in Chinese)

Isolation,Identification and Immunogenicity Evaluation of H9N2 Subtype Avian Influenza Virus HF Strain

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LU Yukui, LIU Jiaqi, ZHONG Jiacheng, CHEN Jidang, ZHU Wanjun, ZHANG Yishan, ZHANG Jipei. Isolation,Identification and Pathogenicity Analysis of Avibacterium paragallinarum from Geese [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2562-2572.
[2]	XIE Shouyu, WANG Ruimin, CUI Pengfei, SHI Kaichuang, WEI Xiankai, FENG Shuping, LONG Feng, QU Sujie, LU Wenjun, HUANG Jinshan, HUANG Fengmei, WEN Xinrui, YIN Yanwen, DENG Guohua. Molecular Characteristics of Complete Genomes of Two H3 Subtype Avian Influenza Virus Isolates Derived from Sea Duck in Guangxi Province [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1319-1328.
[3]	LI Zhijie, ZHAO Ying, MA Xin, WANG Meng, YU Sijiu, WANG Libin, PAN Yangyang. Research Progress on Isolation Methods and Main Reproductive Functions of Exosomes from Animal Follicular Fluid [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1480-1488.
[4]	ZHANG Zhewei, DAI Xiaotong, WEI Zudan, CHEN Yunpeng, GAN Jing, BEI Weicheng. Study on Prokaryotic Expression and Immunogenicity of OmpW and TbpA Proteins in Swine Pasteurella multocida [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1489-1498.
[5]	TIAN Changqing, ZHANG Kunzhong, QI Yumei, SHI Wenjing, DONG Zhijie, ZHANG Haohao, HE Zengwen, ZHI Ji, ZHAO Xuehui, CHONG Qian, XUE Huiwen, GOU Huitian. Isolation,Identification and Biological Characteristics Analysis of a Strain of Listeria monocytogenes from Environment [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1543-1555.
[6]	LIU Jiaxing, HAN Xueying, ZHANG Xinru, DENG Jiaxin, YAO Lunguang, LIU Yangkun. Preparation and Immunogenicity Analysis of Ferritin Nanoparticles Carrying Antigenic Epitopes of Porcine Epidemic Diarrhea Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1567-1574.
[7]	CAO Yingying, LIANG Liang, ZENG Yirong, ZHONG Qinghua, YUAN Xiaofang, TANG Haibo, LENG Jing. Isolation,Identification and Biological Characteristics of Salmonella from Tupaia belangeri chinensis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1718-1728.
[8]	ZHANG Jingxu, XU Yu, LIANG Haiying, ZENG Zhiyong, TANG Deyuan, WANG Bin, XU Songping, WAN Juan, ZHU Yang. Phylogenetic Evolution and Epitope Analysis of Pseudorabies Virus Guizhou Isolates and Vaccine Strains [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1093-1106.
[9]	LIU Kexin, WANG Chao, ZHANG Ao, LIU Jiali, TAN Bin, ZHANG Shuqin. Isolation,Identification and Genome Sequence Analysis of Bovine Viral Diarrhea Virus LN-1 Strain [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1150-1159.
[10]	GAN Lu, ZHENG Huizhen, NUO Mingdalai, LIU Yan, JIN Min, HE Wenwen, WEN Licui, LI Yongchang, GAILIKE·Bayinchahan. Propagation of Trypanosoma evansi Yili Strain and Cloning, Expression and Bioinformatics Analysis of Its PFR Gene [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 469-478.
[11]	WANG Bingyi, MA Hongcai, ZOU Minghao, FAN Shijie, YUAN Zhenjie, Labaciren, DONG Hailong, ZENG Jiangyong. Isolation,Identification and Drug Resistance Analysis of Streptococcus from Yak [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 745-753.
[12]	ZHONG Huachen, WANG Lifang, GUO Chenyang, LIU Jialin, SONG Jie. Isolation,Identification and Drug Resistance Analysis of Pathogenic Bacteria from Milk Samples and Environment of Mastitis in Inner Mongolia [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 817-826.
[13]	SUN Yu, ZHANG Hao, LI Changyang, TAO Yanzi, WANG Qiuju. Isolation,Identification and Biological Characteristics of Antioxidant Bacillus from Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 359-367.
[14]	WEI Jin, WANG Jia, ZHAO Haoran, LIU Bo, LIU Yuanjie, YAO Wensheng, LIU Yan, MA Xin. Isolation, Identification and Antimicrobial Susceptibility of Mycoplasma gallisepticum from Different Areas [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 368-376.
[15]	YANG Fumei, YANG Linfu, ZENG Cheng, YANG Zhi, WANG Kang, DUAN Gang, DAI Feiyan. Isolation,Identification and Biological Characterization Analysis of a Strain of Carnobacterium maltaromaticum from Barking Deer [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 398-407.