以传统方法和杆状病毒表现VP2次单位传染性华氏囊病毒IBDV.DOCVIP

以傳統方法和桿狀病毒表現VP2次單位傳染性華氏囊病毒（IBDV） 蛋白製作IBDV不活化疫苗抗原在雞之免疫保護力比較 Comparison of Immunprotectivity Provided by the Conventional and the Baculovirus Expressied VP2 SubunitProtein of Infectious Bursal Disease Virus （IBDV） as Inactivated IBDV Vaccine Antigen in Chickens 研究生: 許郁停 Yu-Ting, Hsu 指導教授: 林茂勇 Maw-Yeong Lin [摘要] 傳染性華氏囊病病毒（infectious bursal disease virus; IBDV）為造成三至六週齡雞B淋巴芽細胞壞死之嚴重免疫抑制傳染病。IBDV之主要結構蛋白為VP2及VP3，其中VP2具有誘導中和抗體的抗原表面決定位。為生產供目前應用之較具免疫保護力和診斷意義的IBDV-VP2次單位蛋白，以反轉錄-聚合酶連鎖反應增幅具良好交叉免疫保護力之一新台灣野外IBDV分離株（V263/TW02）之VP2全長約1.5 Kbs基因，選殖後定序，比對其基因親源關係後確定其為已發生點變異之超強毒IBDV（vvIBDV）；以V263/TW02 之VP2基因嵌入桿狀病毒表現系統載體可表達出分子量分別為48和21 kDa之A3和C2次單位蛋白，以該IBDV-VP2-C2製成次單位疫苗以與傳統方法製作之同源病毒不活化IBDV疫苗免疫無特定病原雞比較其免疫保護力，結果VP2-C2次單位疫苗之免疫保護力略遜於傳統不活化疫苗，但卻可激發高的酵素連結免疫吸附試驗抗體反應力價。V263/TW02-VP2-C2次單位蛋白或可供作為IBDV之血清學診斷試驗用之抗原，V263/TW02-VP2-A3次單位蛋白之免疫原性尚待進一步試驗探討 關鍵字: 傳染性華氏囊病毒、桿狀病毒表現系統、次單位蛋白、不活化疫苗 【Abstract】 Infectious bursal disease virus (IBDV) is a immunosuppressive infectious disease in causing necrotic of B-lymphoblast in chickens at 3 to 6 weeks of age. Viral protein (VP) 2 and 3 are the mainly important 2 structural proteins of IBDV. The VP2 of IBDV contains the antigenic epitopes in inducing the neutralizing antibody. The whole length of VP2 gene, approximately 1.5 Kb nucleotides, of a recently local isolate (V263/TW02) IBDV being verified with good cross protectivity against several local isolates was amplified with reverse transcriptase-polymerase chain reaction , sequenced and phylogenic analysed of the nucleotide as a very virulent IBDV (vvIBDV) with some point mutation. The gene was then cloned in the vector of the baculovirus expression system for producing the most protective VP2 subunit protein as antigen for IBDV V263/TW02-VP2 subunit vaccine preparation. A3（48 kDa）and C2（21 kDa）2 subunit proteins was found expressied in the system. The V263/TW02-VP2-C2 subunit vaccine was then used to immunized the specific-pathogen-free chickens for comparison of the protectivity with the conventional inactivat