禽流感高免球蛋白的制备及应用研究  

Preparation and Application of Hyperimmune Yolk Ig against Avain Influenza

作　　者：李雁冰[1] 李海燕[1] 辛晓光[1] 田国彬[1] 冯菊艳[1] 唐秀英[1] 于康震[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所动物流感研究中心,黑龙江哈尔滨150001

出　　处：《中国兽药杂志》2002年第9期12-15,共4页Chinese Journal of Veterinary Drug

摘　　要：应用H9N2亚型禽流感病毒(AIV)油乳剂灭活苗6个不同剂量免疫30周龄SPF蛋鸡,通过血凝抑制(HI)和琼脂免疫扩散(AGP)试验监测血清及卵黄抗体消长规律,确定2.0 mL为最适免疫剂量。首免后第18天血清与卵黄中抗体同时升至第一高峰,此时加强免疫。二免后1周,效价达到第二高峰,用常规抽提法从高抗体滴度卵黄中提取高免球蛋白(IgY)。中和试验(NT)表明:IgY具有中和病毒感染的作用,且中和抗体滴度随HI滴度的降低而降低;给鸡肌肉注射后6 h能检测出血清的HI抗体,24 h达到峰值,维持7 d,第13天血清中HI抗体消失。104个EID50的AIV(H9N2)攻毒,同时用HI价为214的IgY的10倍系列稀释物分5组进行保护试验,确定HI价为214的IgY保护范围在100倍稀释与10倍稀释之间。IgY系列保护试验表明HI价28为完全保护的最低滴度;在攻毒的前1天及至攻毒后第5天接种IgY均能100%保护。攻毒保存期试验证明,所制得的高免球蛋白4 ℃可以保存6个月,-20 ℃可保存1年。Thirty-week-old SPF layers were immunized with six different doses of inactivated oil emulsionvaccine against H9N2 Avian Influenza. It was confirmed that the optimal immune dose was 2.0 mL with detectionof the serum and yolk antibody(Ab) titre by hemagglutination-inhibition (HI) and agar gel precipitation (AGP)tests. Both serum and yolk Ab titer peaked 18 days post immunization. After the booster immunization wasgiven, Ab titre reached to another peak. The hyperimmunized yolk Ig (IgY) was extracted by general method.Neutralization test showed that IgY was able to neutralize the virus infection, and NT Ab titer decreased alongwith HI titer. The HI Ab could be detected 6 hours post injected (pi), and peaked 24 hours pi, furthermore holdedline for 7 days and undetected 13 days pi. Layers inoculated with 104 EID50 AIV(H9N2) and injected IgY simulta-neity by serial 10 fold dilution. It was demonstrated that the protection titer was between 26 and 210, and theminimum protection titer was 28. Protection of 100 percent was obtained 6 days pi. IgY remained active after 6months at 4℃ and 1 year at -20℃.

关 键 词：禽流感 高免球蛋白 制备 应用 中和试验 攻毒保护试验 

分 类 号：S858.3[农业科学—临床兽医学] S859.797[农业科学—兽医学]