2014-01-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656449摘要：家禽傳染性支氣管炎病毒 (infectious bronchitis virus, IBV) 對於全世界 及台灣的養雞產業而言是相當重要的呼吸道病原，並且散播速度相當迅速，造成嚴 重的經濟損失。IBV已知血清型種類繁多且不斷出現新的變異株，而台灣本土的病毒 株為獨立演化的病毒，基因型與血清型皆與目前使用的麻州型疫苗株有相當程度的 差異。也因為IBV感染相當普遍且變異株眾多，田間快速診斷試劑的發展日漸受到重 視。本計畫的研究目標即為建立與改善IBV的快速檢測試劑與方法。 首先將生產並篩選抗S1與N蛋白的單株抗體並加以純化，作為後續發展應用；在 抗原檢測方面，目的是利用標示膠體金的單株抗體能特異性的結合不同型別的IBV抗 原，在測試紙片上呈色，發展為免疫色層分析測試紙片，供現場第一線防疫人員及 時得知雞隻感染情形，及早淘汰病雞；在抗體檢測方面，改良先前已初步發展的血 清型別特異性阻斷型ELISA，改以重組病毒蛋白塗鍍，降低成本，取得專利並技轉上 市，供田間樣本大量且快速檢測野外毒感染的抗體存在，並施行野外感染抗體陽性 率調查，作為未來防疫計畫的指標。 103年度計畫預期效益包括產出：a. 抗S1蛋白單株抗體2株 b. 抗N蛋白單株抗 體2株 c. 桿狀病毒表現之S1重組蛋白(除本研究用途外，另可供疫苗測試用途) d. 血清特異性阻斷型ELISA產品技術轉移 e. 期刊論文一篇 f. 研討會論文一篇。 104年度計畫預期效益包括產出：a. 桿狀病毒表現之N重組蛋白(除本研究用途外 ，另可供疫苗測試用途) b. 偵測IBV免疫色層分析測試紙片產品技術轉移 c. 期刊 論文一篇 d. 研討會論文一篇。<br> Abstract: Avian infectious bronchitis virus (IBV) causes tremendous economic loss to the poultry industry worldwide, including Taiwan. IBV is amongst the most important pathogens in chickens and it spreads rapidly in the flocks. In addition to hundreds of known serotypes, new viral variants have emerged due to viral evolution and antigenic variation in IBVs. Therefore, the development of a sensitive, specific, and easily performed assay is crucial for the rapid detection and surveillance of IBV infections. The aim of this project is to develop rapid detection methods against IBV antigen and antibody. Firstly, monoclonal antibodies (mAbs) against the S1 and N protein of IBV will be produced and purified for further application. For antigen detection, the selected mAbs will be labeled with colloidal gold and visible color will appear on the strip when different genotypes of viruses are captured by the detecting mAb. For antibody differentiation, the existing serotype-specific blocking ELISA will be validated and improved by replacing the coating antigen with the IBV S1 recombinant protein. Sensitivity, specificity, and the limit of detection of these two detection methods will be carefully evaluated. Antibody prevalence of IBV will be surveyed. The final goals of this project are to commercialize the rapid test products for the use in the field. During the first year of the project, the following items are expected: two anti-S1 mAbs, two anti-N mAbs, baculovirus-expressed S1 recombinant protein, commercialized blocking ELISA Kit, and journal/conference articles. For the second year of the project, the following items are expected: baculovirus-expressed N recombinant protein, commercialized immunochromatographic strip test, and journal/conference articles.家禽傳染性支氣管炎病毒單株抗體免疫色層分析試紙片Avian infectious bronchitis virusMonoclonal antibodyImmunochromatographic strip test