Standardization of an enzyme-linked immunosorbent assay for detection of antibody to avian reticuloendotheliosis virus
2005
Sung, H.W. (Kangwon National University, Chunchon, Republic of Korea), E-mail: sunghw@kangwon.ac.kr | Lee, S.J. (Kangwon National University, Chunchon, Republic of Korea)
Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the 107,000×g for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen (1 ㎍/well) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum).
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