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

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 Xiang-rui LI Yin 

作者机构：Institute of Veterinary MedicineJiangsu Academy of Agricutural Sciences/Key Laboratory of Veterinary Biological Engineering and TechnologyMinistry of Agriculture/National Center for Engineering Research of Veterinary Bio-Products College of Veterinary MedicineNanjing Agricultural University 

出 版 物：《Journal of Integrative Agriculture》 (农业科学学报（英文版）)

年 卷 期：2013年第12卷第9期

页      面：1638-1643页

核心收录：

学科分类：0710[理学-生物学] 0832[工学-食品科学与工程（可授工学、农学学位）] 0830[工学-环境科学与工程（可授工学、理学、农学学位）] 1004[医学-公共卫生与预防医学(可授医学、理学学位)] 1001[医学-基础医学(可授医学、理学学位)] 0905[农学-畜牧学] 0906[农学-兽医学] 100102[医学-免疫学] 0901[农学-作物学] 0703[理学-化学] 0902[农学-园艺学] 10[医学] 0713[理学-生态学] 

基　　金：supported by the National Natural Science Foundation of China (31172345) the Jiangsu Provincial Agricultural Science and Technology InnovationFoundation, China (cx(11)4039) 

主　　题：goose flavivirus double antibody sandwich ELISA monoclonal antibodies 

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.m）. The geese flavivirus could be detected at a minimal concentration of 1.875 μg 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.