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題 名 | Detection of Antibody against Avian Leukosis Virus Subgroup A by Enzyme-linked Immunosorbent Assay with Baculovirus Expressed gp85 Protein=利用桿狀病毒表現家禽白血病A亞群gp85蛋白質以酵素連結免疫吸附法檢測抗體 |
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作 者 | 許萌芳; 王金和; 萬灼華; | 書刊名 | 臺灣獸醫學雜誌 |
卷 期 | 40B:S1 2014.03[民103.03] |
頁 次 | 頁39-45 |
分類號 | 437.245 |
關鍵詞 | 抗體的檢測; 家禽白血病病毒; 桿狀病毒載體; 酵素免疫連結吸附法; A亞群; Antibody detection; Avain leukosis virus; Baculovirus vector; Enzyme-linked immunosorbent assay; Subgroup A; |
語 文 | 英文(English) |
中文摘要 | 為了檢測家禽白血病A亞群病毒,(avian leukosis virus subgroup A, ALV-A) 的抗體,選殖台灣分離ALV-A 的gp85基因,利用桿狀病毒載體表現系統進行ALV-A 之表面醣蛋白gp85表現。重組桿狀病感染後之昆蟲細胞, 進行西方墨點法分析及免疫螢光染色法可偵測到重組gp85蛋白質之表現。重組gp85蛋白質直接做為塗鍍抗原,與 標準陽性和陰性血清進行ELISA測試,確認ALV-A抗血清可造成ELISA讀值上升,SPF雞隻血清則否。對田間樣本 血清進行測試,以血清中和試驗做為抗體檢測為金標準,得到ELISA 之敏感性為80% (24/30),特異性為83.3% (35/42)。因此利用桿狀病表現系統進行之ALV-A gp85蛋白表現及酵素連結免疫吸附法可應用於現場抗ALV-A 抗 體的檢測。 |
英文摘要 | To detect antibody against avian leukosis virus subgroup A (ALV-A), the gp85 gene of an ALV-A strain isolated in Taiwan was cloned and expressed in a baculovirus expression vector. The expressed protein confirmed by Western blotting and immunofluorescence tests with anti-ALVA antiserum was coated in 96-well plates and was used as an antigen for antibody detection by an indirect enzyme-linked immunosorbent assay (ELISA). Neutralization test performed in chicken sera from the infected and uninfected flocks was used as the standard to evaluate the accuracy of the ELISA. The cut-off point was calculated by receiver operator characteristic (ROC) curve to be 1. The sensitivity and specificity of this ELISA were 80% (24/30) and 83.3% (35/42), respectively. In conclusion, this ELISA could be applied for the anti-ALV-A antibody detection in the field. |
本系統中英文摘要資訊取自各篇刊載內容。