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標題: 用家禽里奧病毒蛋白σNS單源抗體為媒介開發捕捉型-ELISA之診斷試劑
Development of a Monoclonal Antibody Capture-Elisa for the Diagnosis of Avian Reovirus Infection
作者: 李龍湖
關鍵字: 應用研究
生物技術, 畜牧獸醫類
摘要: 本計畫擬將融合瘤細胞打入小白鼠腹腔生產單源抗體.腹水純化後, 用以塗度ELISA測定盤.接著以pET32a表現系統表現病毒σNS, 打破的細菌上清液加入含單源抗體孔洞中.σNS將為抗體吸附, 如此用以測定SPF陽性及陰性血清.此外, 疫苗用不同方式免疫雞隻.每週採血, 收集血清, 再用前述ELISA盤進行抗體檢測.該研發之ELISA亦將用於野外收集血液抗體檢測.同時, 以血液中和試驗, 檢測血清病毒中和抗體.所得結果將和ELISA之結果比較, 並分析其抗體消長之關連性.
This research proposal is produce monoclonal antibodies by injecting, eitra peritoneally, hybridoma cells into BALB/c mice which have been previously primed prestane.primed with prestane.Following antibody purification from ascitic fluids collected form mice, the antibodies are coated on ELISA plates.Subsequently, the bacterial cell lysates containing σNS protein synthesized in E.coli containing pET32a-σNS are added into each well of plates.The protein σNS is captured by antibodies immobilized on the plate, then is used to detect antibodies-positive and-negative sera from SPF chickens.In addition, avian reovirus vaccine are used to immunize chicks with various routes.The sera are collected weekly and the antibody is tested by using the ELISA as mentined above.The ELISA described have is also used to detect the sera collected from the chicken flocks.Serum neutralization test is conducted to detect the neutralizing antibodies of the collected sera.The results from serum neutralization test and from ELISA are compared.The correlation rates between neutralization antibody titer and the OD value of ELISA are determined.
其他識別: 90農科-6.2.1-檢-B5(6)
Appears in Collections:獸醫學系所



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