Development of quickly diagnosis for silkworm virus by use of monoclonal antibody

为了建立一种简便、快速的家蚕病毒诊断方法，我们采用杂交瘤筛选法制备了抗家蚕细胞质型多角体病毒（CPV）和抗家蚕核型多角体病毒感染增强因子（EF）的单克隆抗体。其中一组对CPV-I株有显著反应，第二组对CPV-H株有反应，第三组对CPV-H株有中度反应。结果表明，两株病毒均具有CPV-H和CPV-I的共同抗原表位。但有些菌株的反应在两个菌株之间略有不同.粘虫痘病毒产生的包涵体有两种类型。增强因子（Enhancing Factor，EF）是一种能增强核型多角体病毒感染力的蛋白质，因此我们研制了针对增强因子的单克隆抗体。但这个莫 ...更多信息 非克隆抗体对球状体的主要结构蛋白spheroidin无反应。而另一种包涵体蛋白纺锤体则与该单克隆抗体反应显著。结果表明，纺锤体的主要结构蛋白49.5KDa蛋白与增强因子具有共同的抗原表位，本研究尝试用单克隆抗体和晶体双功能酶构建免疫传感器，以简便、快速地检测家蚕细胞质多角体病毒。我们首先从抗犬细小病毒单克隆抗体中纯化IgG，并对其反应性进行了检测。结果表明，该单克隆抗体具有较高的效价，用ELISA法测定，1 × 10 ~（-1）μ <-5>g IgG可检出1 μ g/ml的CPV，用Westernblotting分析表明，该单克隆抗体能与CPV核心蛋白和多角体蛋白反应。用蛋白A涂敷的晶体双抗体偶联的IgG呈同频变化。通过去除非特异性吸附，可以研制出高灵敏度的免疫传感器。少

In order to develop a simple ana quickly diagnosis for silkworm virus we made the monoclonal antibodies for cytoplasmic-polyhedrosis virus (CPV) and dnhancing factor of nuclear-polyhedrosis virus infection (EF) by hybridoma selection method.1.Three-group clones of monoclonal antibody for CPV-I strain were obtained. Thst is, one group was remarkably reacted to CPV-I strain, the second reacted to CPV-H strain, and the third reacted in the intermediate degrees. These results suggested that both strains pessessed the common epitope between CPV-H and CPV-I strains. But some of them showed a little different reaction between two strains.2. There are two types of inclusion body produced by Pseudaletia separata entomopoxvirus. One is called spheroid enclosed virus particles and the other is spindle without virus particle in it. Enhancing factor (EF) is known as the increasing the nuclear-polyhedrosis virus infection.So we mede a monoclonal antibody for the enhancing factor protein. But this mo … More noclonal antibody did not react for spheroidin which was a main constructed protein of spheroid. On the contrary the another inclusion protein, spindle, reacted remarkably to this monoclonal antibody. From the result it was demonstrated that 49.5KDa protein, i.e.main constructed protein of spindle, had common epitope to enhancing factor.We tried to develop the immunosensor constructed by monoclonal antibody and crystal bibrator to detect the cytoplasmic-polyhedrosis virus of the silkworm simply and quickly. At first we purified the IgG from monoclonal antibody for CPV and tested its reaction. The IgG showed the highly titers, i.e.1 x 10^<-5> g IgG could detect 1 ug/ml CPV by ELISA.In addition to this it was clear that this monoclonal antibody reacted both CPV core protein and CPV polyhedrin by western blotting analysis. The IgG conjugated the crystal bibrator smeared with protein A was showed the variation of co-frequency. By remove the non-specific adsorption we might develop the high sensitive immunosensor. Less