[1]徐海峰,张 莹,刘 琳,等.兼顾H5亚型高致病性禽流感病毒不同分支HA蛋白单克隆抗体的制备及鉴定[J].中国预防兽医学报,2018,(07):639-642.[doi:0.3969/j.issn.1008-0589.201709019]
 XU Hai-feng,ZHANG Ying,LIU Ling,et al.Preparation and identification of monoclonal antibodies against HA protein from different H5 subtypes of highly pathogenic avian influenza virus[J].Chinese journal of preventive veterinary medicine,2018,(07):639-642.[doi:0.3969/j.issn.1008-0589.201709019]
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兼顾H5亚型高致病性禽流感病毒不同分支HA蛋白单克隆抗体的制备及鉴定()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年07
页码:
639-642
栏目:
研究简报
出版日期:
2018-08-15

文章信息/Info

Title:
Preparation and identification of monoclonal antibodies against HA protein from different H5 subtypes of highly pathogenic avian influenza virus
文章编号:
1008-0589(2018)7-0639-04
作者:
 

徐海峰1张 莹1刘 琳1曾显营1谷琳琳1熊晓宇2王秀荣1*陈化兰1

 (1. 中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室,黑龙江 哈尔滨 150069;
2. 东北农业大学 动物医学学院,黑龙江 哈尔滨 150030)
Author(s):
 

XU Hai-feng1 ZHANG Ying1 LIU Ling1 ZENG Xian-ying1 GU Lin-lin1 XIONG Xiao-yu2WANG Xiu-rong1* CHEN Hua-lan1

 (1. State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences,
Harbin 150069, China; 2. College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China)
关键词:
H5亚型禽流感病毒基因型分支单克隆抗体
Keywords:
  H5 subtype avian influenza virus  genotype branch  monoclonal antibody
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201709019
文献标志码:
A
摘要:
为建立针对我国主要流行的H5亚型Clade7.2、Clade 2.3.2和Clade2.3.4.4分支高致病性禽流感病毒(HPAIV)不同流行株的免疫学快速检测方法,本研究将Clade7.2分支禽流感疫苗株CK/LN/S4092/2011(H5N1)(Re-7)、Clade2.3.4.4分支疫苗株DK/GZ/4/2013(H5N1) (Re-8)和Clade2.3.2.1e分支疫苗株DK/AH/S1246/14(H5N1)(Re-10)混合作为免疫原免疫4周龄BALB/c雌鼠,4次免疫后无菌取其脾细胞和骨髓瘤细胞(SP2/0)在聚乙二醇促融剂的作用下融合,通过间接ELISA方法及HI试验筛选获得6株能稳定分泌抗血凝素蛋白(HA)的单克隆抗体(MAb)杂交瘤细胞,分别命名为1B10、2A10、2C8、3E6、4G9和H2B1。经检测1B10、2A10、2C8和H2B1能够与当前AIV流行的分支反应,具有较好的广谱性。本研究为H5亚型AIV快速诊断技术的研发提供基础材料。
 
Abstract:
To establish a rapid detection method against HA protein of Clade7.2, Clade 2.3.2 and Clade2.3.4.4 H5 subtype avian influenza virus (AIV), which are mainly prevalent in China, the Clade7.2 branch vaccine strain CK/LN/S4092/2011(H5N1)(Re-7), Clade2.3.4.4 branch vaccine strain DK/GZ/4/2013(H5N1)(Re-8) and Clade2.3.2.1e branch vaccine strain DK/AH/ S1246/14(H5N1) (Re-10) were used as an immunogen to immunize four-week old BALB/c female mice. After four immunizations, the splenic cells of mice were fused with SP2/0 cells. Then six hybridoma cell lines which stably secreting the monoclonal antibodies (MAb) against HA protein were screened by indirect ELISA and HI assay, named 1B10, 2A10, 2C8, 3E6, 4G9 and H2B1. The MAb of 1B10, 2A10, 2C8 and H2B1 were able to react with the different recently prevalent H5 Clade AIVs, and they had a broad spectrum. This study provides basic materials for the rapid diagnosis of H5 subtype AIV.

参考文献/References:

[1]Yu Zhi-jun, Gao Xiao-long, Wang Tie-cheng, et al. Fatal H5N6 avian influenza virus infection in a domestic cat and wild birds in China [J]. Sci Rep, 2015, 5: 10704.
[2]WHO(2017). Cumulative number of confirmed human cases for avian influenza A(H5N1) reported to WHO [EB/CD]. http://www.who.int/influenza/human_animal_interface/2017-07-25-
Table H5N1, 2017.
[3]Xu Xi-yan, Kanta S, Cox N J, et al. Genetic characterization of the pathogenic influenza A/Goose/Guangdong/1/96 (H5N1) virus: similarity of its hemagglutinin gene to those of H5N1 viruses from the 1997 outbreaks in Hong Kong [J]. Virology, 1999, 261(1):15-19.
[4]WHO/OIE/FAO H5N1 Evolution Working Group. Toward a unified nomenclature system for highly pathogenic avian influenza virus (H5N1) [J]. Emerg Infect Dis, 2008, 14(7): e1.
[5]Sun Hong-lei, Sun Yi-Peng, Pu Juan et al. Comparative virus replication and host innate responses in human cells infected with three prevalent clades (2.3.4, 2.3.2, and 7) of highly pathogenic avian influenza H5N1 viruses [J]. J Virol, 2014, 88 (1): 725-729.
[6]Crowther J R.The ELISA guidebook [M]. USA: Humana Press, 2009.
                       (本文编辑:李 娜)

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更新日期/Last Update: 2018-08-15