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標題: The Efficacy and Commercialization of Recombinant Atrophic Rhinitis Subunit Vaccine
作者: 簡茂盛
關鍵字: 生物技術, 畜牧獸醫類
摘要: 豬萎縮性鼻炎為感染豬隻之重要細菌性疾病, 而本病於台灣豬場之感染率高達75%, 目前國外進口之商品化AR疫苗因生產PMT類毒素步驟繁瑣, 且病原在自然培養狀況下毒素產量又極低, 故PMT類毒素疫苗價格十分高昂, 若疫苗中只添加不活化之菌體以取代類毒素時, 其免疫效力又極端不足, 而所萃取之細胞毒素又有不活化完全時其安全性之慮, 導致農民使用AR疫苗意願低落.本實驗室曾對AR作一系列之研究, 同時為得到高產量與安全性之PMT毒素, 因此以基因工程技術研發次單位PMT毒素疫苗, 基因重組次單位疫苗則有別於傳統的疫苗, 其優點包括一旦製備完成後, 生產過程簡便且容易大量製造, 故成本遠低於傳統疫苗.重組次單位疫苗亦能呈現大量不具毒性之抗原蛋白, 誘發母豬產生高力價抗體, 並藉由移行抗體保護仔豬在初生1-3週高危險期間的防禦能力, 此外, 仔豬初生後經2次重組次單位疫苗單獨或經混合不活化A型P.multocida, 對豬隻皆具有良好的保護能力, 亦可測得具保護性之中和抗體力價, 因此重組次單位類毒素疫苗成為防治AR最具潛力的疫苗種類.本實驗計畫的目的即應用所研發完成之AR重組PMT次單位疫苗, 進行技術轉移給國內疫苗廠商, 並評估其所製造的新一代AR重組次單位PMT疫苗, 於田間豬場在母豬與仔豬的免疫保護效力與抗體消長之情形, 期望藉由本實驗能成功移轉高效能且具國際經濟競爭性的重組次單位AR疫苗, 以貢獻國內養豬業者.
The toxin-producing strains of Pasteurella multocida ( both type D and A )are characterized to be the major causative agent of atrophic rhinitis ( AR )in swine.The P.multocida toxin ( PMT )with a calculated molecular weight of 146kDa has also been identified to cause the atrophy of nasal turbinate and skeletal bones.In our laboratory, the full length and partial of PMT gene were cloned in pET-system expression vectors and were applied for developing the subunit PMT vaccine.Several clones contained partial or complete open reading frame of 3.8kb in size of PMT gene were successfully constructed into different vectors.The products of different clones could be identified both with polyclonal and monoclonal antibody by Western blotting assay.The expressed of PMT fusion proteins can also mount good protective immunity in experimental swine when they were challenged with high dosage of PMT toxin.Furthermore, the neutralizing antibody titer was markedly increased with vero cell toxicity test.After twice vaccination, the sows mount good maternal antibodies to protect newborn from AR formation.The purpose of this project is to provide the tested subunit clones for AR vaccine trial and transfer the techniques for local company in order to control disease and decrease the economic loss in pig farm.
其他識別: 90農科-2.1.3-檢-B3
Appears in Collections:獸醫病理生物學所



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