草莓伪温和黄边病毒抗血清制备及血清学检测技术研究

Studies on the Preparations of Antiserum and Serological Detection Techniques for Strawberry Pseudo Mild Yellow Edge Virus

将提纯的草莓伪温和黄边病毒（ＳｔｒａｗｂｅｒｒｙｐｓｅｕｄｏｍｉｌｄｙｅｌｌｏｗｅｄｇｅｖｉｒｕｓＳＰＭＹＥＶ）作抗原免疫家兔，获得效价较高的抗血清，经ＳＤＳ－对流免疫电泳测其效价为１：１２８．四点疫吸附固定法检测ＳＰＭＹＥＶ效果甚理想，可检测到微量病素，提纯病毒浓度低至０．０５μｇ／ｍｌ仍有效果．酶联免疫吸附测定法检出灵敏度也相当高，当病毒浓度为０．１μｇ／ｍｌ时及病叶汁液稀释至１０２４倍时，仍呈阳性反应．免疫电镜技术检视，ＳＰＭＹＥＶ抗血清能清晰地”捕获”同源病毒，并且有装饰作用，抗血清以稀释为１：５００时“捕获”病毒粒子最多．

High titer preparations of atiserum was obtained by using purified SPMYEV to immunize rabbitis from which anti-SPMYEV IgG was also purified by ammonium sulfate precipitation, DE32 columm chromatograhy. Dot immunobinding assay (DIBA) Indirect enzyme linked immunosorhent assay (ELIST) and Immunosorbent electron microscopy (ISEM) technique were used and compared for virus identification. According to the experimental data it showed that the DIBA is sensitive to detect the purified SPMYEV Of 0.05 μg/ml. while the sensitivity of indirect ELISA showed 0.10μg/ml. Diseased leaf sap diluted in 1:1024 could also be use effetively for abvove mentioned methods. In ISEM test, a lot of homologous virus particles (SPMYEV) could be trapped on the grids which were coated with antiserum against SPMREV ,but heterogous virus particles were not. The sensitivity and soecificity of ISEM dipended on the concentration of the antiserum acted acted in the grid The results showed that the optimal dilution of antiserum was 1:500.ISEM also showed very good effctivity for virus identiffcation