基于血凝素蛋白特异性抗体的H7亚型流感病毒双抗体夹心ELISA检测方法建立  

Establishment of a double antibody sandwich ELISA for detection of H7 subtype influenza virus with hemagglutinin specific antibodies

作　　者：王泽敏 祁贤[2] 鲍倡俊[2] 焦永军[2] 卫平民[1] WANG Ze-min;QI Xian;BAO Chang-jun;JIAO Yong-jun;WEI Ping-min(School of Public Health,Southeast University,Nanjing 210009,China;National Health Committee Key Laboratory of Enteric Pathogenic Microbiology,Jiangsu Provincial Center for Disease Control and Prevention,Nanjing 210009,China)

机构地区：[1]东南大学公共卫生学院,南京210009 [2]江苏省疾病预防控制中心,国家卫生健康委员会肠道病原微生物重点实验室,南京210009

出　　处：《中国人兽共患病学报》2023年第5期455-460,共6页Chinese Journal of Zoonoses

基　　金：南京市重大科技专项(No.2021-11005)。

摘　　要：目的基于H7亚型流感病毒,制备针对于HA抗原的单克隆抗体,并建立双抗体夹心ELISA检测体系,为其在流感病毒快速检测试剂研发中提供技术储备。方法用H7亚型禽流感病毒HA重组蛋白免疫BALB/c小鼠,通过传统杂交瘤技术制备HA特异性单克隆抗体,筛选可在双抗体夹心ELISA技术平台上使用的配对抗体,初步探索该ELISA体系的检测特异性、敏感性和广谱性。结果共制备出6株HA特异性单克隆抗体,并成功筛选出捕获抗体8B11-C9及检测抗体7C3-G5的最佳配对。该双抗体夹心体系对HA重组蛋白的检测灵敏度可达到1.56 ng/mL,能有效检测出不同宿主来源的H7亚型流感病毒,与其他亚型流感病毒无交叉。结论成功制备出H7亚型流感病毒HA单克隆抗体,建立了H7亚型流感病毒特异性的双抗体夹心ELISA检测方法,为其快速检测试剂研发奠定了基础。Monoclonal antibodies against the HA antigen of H7 subtype influenza virus were prepared,and a double antibody sandwich ELISA system was established to support research and development of rapid detection reagents for influenza virus.The H7 subtype avian influenza virus HA recombinant protein was used to immunize BALB/c mice,and HA-specific monoclonal antibodies were prepared with a conventional hybridoma technique to screen for paired antibodies that could be used in a double antibody sandwich ELISA platform.The specificity,sensitivity and broad spectrum of the ELISA system were preliminarily explored.Six HA-specific monoclonal antibodies were prepared,and the optimal pairing of the capture antibody 8B11-C9 and detection antibody 7C3-G5 was selected.The double antibody sandwich system detected the recombinant HA protein with a sensitivity of 1.56 ng/mL and effectively detected H7 subtypes of influenza viruses with different host origins without cross-reactivity with other subtypes.The monoclonal antibody to H7 subtype of influenza virus HA was successfully prepared,and a double antibody sandwich ELISA for H7 subtype influenza virus was established,thus laying a foundation for development of future rapid detection reagents.

关 键 词：H7亚型流感病毒 单克隆抗体 HA蛋白 双抗体夹心ELISA 

分 类 号：R373.1[医药卫生—病原生物学]