小西葫芦花叶病毒(ZYMV)抗血清制备及检测技术研究  

Study on the Antiserum Preparation and Detection Technology of Zucchini Yellow Mosaic Virus(ZYMV)

作　　者：程珉[1] 张大伟[1] 范卫红[1] 王杰[1] 

机构地区：[1]安徽农业大学植保学院,安徽合肥230036

出　　处：《安徽农业科学》2005年第5期805-806,共2页Journal of Anhui Agricultural Sciences

基　　金：安徽省自然科学基金项目(01011403)资助

摘　　要：从小西葫芦黄化花叶病毒(ZYMV)免疫的母鸡蛋卵黄和兔抗血清中分别提取免疫球蛋白IgY和IgG,用多种血清学方法进行检测。结果表明,免疫鸡和兔25d后,鸡蛋卵黄IgY和兔抗血清的效价分别达到1/64和1/512。在琼脂扩散试验中,当抗体稀释28倍时,与抗原反应无明显沉淀线。在胶乳凝集试验中,抗体稀释210倍时,与抗原反应无明显凝集物。ELISA法以辣根过氧化酶标记抗体IgY和IgG,用双抗夹心法成功地检测出了ZYMV,当抗体稀释106倍时,与抗原反应仍呈阳性。免疫电镜方法检测ZYMV,较一般血清学方法可多捕捉50倍的病毒颗粒。In this paper, immunoglobulin Y and immunoglobulin G were extracted from the yolk of hens and rabbits, which were immunized with Zucchini yellow mosaic virus(ZYMV). The IgY and IgG were etected by means of several kinds of serological reaction and the results were compared. The results showed: immunized after twenty-five days, the titer of antiserum of IgY and IgG by means of tube precipitation were 1/64 ,1/512, respectively. In agar diffusion test, there was no clear precipitin bands formation when antibodies were diluted to 28. In agglutination test, there was no apparent agglutinant when antibodies were diluted to 210. IgY and IgG marked with horseradish peroxidase and ZYMV were detected successfully by means of the double-antibody sandwich method of direct ELISA. It reached a high sensitivity. The antibody reacted positively with antigen when the antibody was diluted to 106. Immunosorbent electron microscopy(ISEM) could detect ZYMV, and it could trap fifty times by this kind of method than by normal serology methods.

关 键 词：小西葫芦花叶病毒 抗血清 制备技术 检测技术 葫芦科作物病害 琼脂扩散法 胶乳凝集反应 免疫电镜 酶联免疫吸附试验 

分 类 号：S436.42[农业科学—农业昆虫与害虫防治]