【摘要】 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。更多还原

【Abstract】 Soybean mosaic virus(SMV)was purified from infected leaf tissue,The procedures of purification involved precipitation with PEG(M.W.6000),two differential centrifugation cycles and 10-40% sucrose density-gradient centrifugation.Size of the virus particles was 714 nm in lengthand 12 nm in diameter.Purified SMV was used to immunize rabbits for the preparation ofantiserum,from which Anti-SMV IgG was purified by ammonium sulfateprecipitation,DE32 column chromatography.The horseradish peroxidase(RZ 3.0)was used to prepare conjugated IgG.In ELISA,the coatingIgG concentration is 4μg/ml and the conjugated IgG concentration is4μg/ml.Our data showed that the sensitivity of ELISA to detect SMV inseed reached a titre of 1:2208 and in diseased leaf sap of 1:1536,whilethe sensitivity of double diffusion test is only 1:28.The virus concentration in germs and radicles was higher than incotyledons and tests.更多还原