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Application on competitive enzyme-linked immunosorbent assay for detecting antibodies to avian reovirus
|關鍵字:||avian reovirus;家禽里奧病毒;competitive ELISA;antibody detection;競爭性酵素接合免疫吸附法;抗體檢測||出版社:||獸醫學系||摘要:||
A competitive enzyme-linked immunosorbent assay (c-ELISA) was developed for detection of avian reovirus (ARV) antibodies. Cell lysates of chicken embryo fibroblast (CEF) infected with ARV (S1133 strain) was used as the coating antigen, and ARV protein-specific monoclonal antibodies (MAb(23), MAb(81), MAb(7), MAb(H1E1) and MAb(4B9)) as competitive factors in the c-ELISA. Sera of several groups of SPF chickens with different vaccination programs or infected with S1133 in different routes were collected and antibody against ARV proteins was analyzed by c-ELISA and virus neutralization test. The OD value of c-ELISA was transformed to percentage inhibition (PI), and the results showed that the PI of antibodies raised up to 40-60 % in 2 weeks postinoculation with inactivated vaccine. Instead, when chickens were inoculated with attenuated live vaccine or infected once with S1133, the PI of antibodies had no significant raise (< 10%). The PI of antibodies against structural proteins raised to 40-60%, and that against nonstructural proteins raised to 10-50%. Besides, antibodies against different viral proteins seemed to appear at the same time. In general, the antibody titers were variable, and some obvious positive responses were of short duration. In virus neutralization test, chickens primed with attenuated and boosted with inactivated vaccine induced more neutralizing antibody production. Compared with c-ELISA titers, the neutralizing antibody titers started to raise in 3 weeks postinoculation and seemed to have lower sensitivity. Using c-ELISA to analyse antibodies of field samples, the results were different from those tested by commercial ELISA kit. However, this c-ELISA was sensitive in detecting ARV-specific antibodies in chicken sera, and was more advantageous than virus neutralization test. The c-ELISA would be an efficient test for ARV serological analysis.
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