Development of Double Antibody Sandwich ELISA for Detection of Duck or Goose Flavivirus
Abstract
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 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.
Date
01-09-2013Author
Hui-min NIU
Xin-mei HUANG
Kai-kai HAN
Yu-zhuo LIU
Dong-min ZHAO
Jing-feng ZHANG
Fei LIU
Tong-tong LI
Xiao-bo ZHOU
Xiang-rui LI
Yin LI
Metadata
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http://www.sciencedirect.com/science/article/pii/S2095311913603329http://digilib.fisipol.ugm.ac.id/repo/handle/15717717/41006
