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大豆花叶病毒(Soybean mosaic virus 简称SMV),经聚乙二醇沉淀、差速离心、蔗糖密度梯度离心提纯后免疫家兔。得到效价为512—1024的抗血清。免疫球蛋白IgG 以饱和硫酸铵沉淀和DE32柱层析提纯,用过碘酸盐氧化法制备标记抗体。ELISA 测定方法为双抗体夹心法。实验结果表明,用ELISA 测感染SMV 的大豆叶片粗汁液的最高稀释度为1536。能测定的单粒种子的最高稀释度为2208,而双扩散能测定的最高稀释度仅28。测定发芽种子不同部位的结果是胚芽和胚根的含毒量高于子叶和皮的含毒量。由北京地区的SMV 抗血清制备的标记抗体同样能用于南京、黑龙江、武汉、辽宁、山东等地区SMV 毒源的测定。
Soybean mosaic virus (SMV) was used to immunize rabbits by polyethylene glycol precipitation, differential centrifugation and sucrose density gradient centrifugation. An antiserum with a titer of 512-1024 was obtained. Immunoglobulin IgG was purified by saturated ammonium sulfate precipitation and DE 32 column chromatography, and the labeled antibody was prepared by periodate oxidation. ELISA method for the double antibody sandwich method. The experimental results showed that the maximum dilution of the crude soybean leaf juice that was infected with SMV by ELISA was 1536. The maximum dilution of a single seed that can be measured is 2208, while the maximum dilution determined by double diffusion is only 28. Determination of germinating seeds in different parts of the results of the germ and radicle toxic higher than cotyledons and skin toxic. The labeled antibody prepared by SMV antiserum in Beijing could also be used for the determination of SMV virulence in Nanjing, Heilongjiang, Wuhan, Liaoning and Shandong.