Development of Double Antibody Sandwich ELISA for Detection of Duck or Goose Flavivirus

文献类型: 外文期刊

第一作者: Niu Hui-min

作者: Niu Hui-min;Huang Xin-mei;Han Kai-kai;Liu Yu-zhuo;Zhao Dong-min;Zhang Jing-feng;Liu Fei;Li Tong-tong;Zhou Xiao-bo;Li Yin;Niu Hui-min;Liu Fei;Li Tong-tong;Zhou Xiao-bo;Li Xiang-rui

作者机构:

关键词: goose;flavivirus;double antibody sandwich ELISA;monoclonal antibodies

期刊名称:JOURNAL OF INTEGRATIVE AGRICULTURE ( 影响因子:2.848; 五年影响因子:2.979 )

ISSN: 2095-3119

年卷期: 2013 年 12 卷 9 期

页码:

收录情况: SCI

摘要: In order to establish double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for detection of duck or goose flavivirus, polyclonal antibody against the flavivirus strain JS804 in geese and monoclonal antibody against the E protein of flavivirus strain JS804 in geese were used as the capture antibody and detection antibody, respectively. The optimal dilution of the capture antibody and detecting antibody capable of detecting the flavivirus strain JS804 in geese were 1:3 200 and 1:160 in the check-board titration, respectively. The reaction time of sample was 1 h, and the optimal working dilution of HRP-labeled goat-anti-mouse IgG was 1:10 000. The positive standard value was 0.247 (OD450 nm). The geese flavivirus could be detected at a minimal concentration of 1.875 mu g mL(-1). The ELISA had no cross-reaction with Newcastle disease virus (NDV), Avian influenza virus (AIV), Infectious bronchitis virus (IBV), Infectious bursal disease virus (IBDV), Duck hepatitis virus (DHV), and Gosling plague virus (GPV). Twenty clinical samples were detected by the DAS-ELISA and RT-PCR respectively, with the agreement rate of 75%. The results revealed that the DAS-ELISA possessed favorable specificity and higher sensitivity, indicating a suitable method for rapid detection of the duck or goose flavivirus.

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