增進豬流行性下痢病毒(PEDV)疫苗效力之研究

豬流行性下痢 (PED)乃是全球養豬業界危害非常嚴重的消化道傳染疾病，尤其是造成哺乳仔豬極為嚴峻的損失。台灣在2014年爆發PED，至今仍影響台灣的養豬產業。傳統的疫苗免疫方式無法讓初生乳豬產生足夠免疫力，因此本研究將針對誘發黏膜免疫反應的口服疫苗進行開發，利用多種油質測試作為口服疫苗之佐劑。而最終研究採月見草油及角鯊烯油作為口服疫苗之油質佐劑；另則以SPAN85與Tween80當介面活性劑並在適當的HLB (Hydrophilic and lipophilic balance)值以及合適的SOR (Surfactant oil ratio)比例下配製70%的油相佐劑加30%的水相病毒抗原之口服疫苗。利用滴水測試，離心動作，顯微鏡鏡檢、之後放置於4度C，室溫，37度C三種狀態，進行穩定觀察4周以上，最終再以人工胃液來作測試進行分析。所得配方混和維他命A及不活化病毒進行小鼠動物實驗。實驗結果中，我們利用總IgG、IgA、細胞增生實驗及中和抗體分析檢視佐劑配方是否能夠達到預期的保護效果，並且製作一支有效之口服油質佐劑不活化疫苗。

關鍵字

PEDV ；不活化疫苗；油質佐劑；維他命A

並列摘要

Porcine epidemic diarrhea (PED) is a very serious infectious disease in the global pig industry. This disease has caused high mortality to suckling piglets. The outbreak of PED in Taiwan in 2014 still affected Taiwan's pig industry. Traditional immunization methods cannot produce sufficient immunity for newborn suckling pigs. Therefore, this study will aim to develop oral vaccines that induce mucosal immune responses against PED. We have tested various oil as adjuvants for developing oral vaccines. In the final study, evening primrose oil and squalene oil were used as the oil-based adjuvants for oral vaccines; SPAN85 and Tween80 were used as surfactants, and 70% oil-phase adjuvants were formulated under appropriate HLB (Hydrophilic and lipophilic balance) values and appropriate SOR (Surfactant oil ratio) ratios. An oral vaccine containing 30% aqueous viral antigen. After the formulation, the oil adjuvants were analyzed by dripping test, centrifugation, microscopic examination. Then the oil adjuvants were storage at 4°C, room temperature or 37°C observation for more than 3 months. After the storage time, the adjuvant was tested in artificial gastric juice. The obtained formulations were mixed with vitamin A and inactivated virus for oral vaccine in mouse animal experiments. In the experimental results, we use total IgG, IgA, cell proliferation assay and neutralizing antibody analysis to check whether the adjuvant formula can achieve the expected protective effect, and make an effective oral oil-based adjuvant inactivated vaccine.