应用斑点法检测植物病毒的研究

应用斑点法检测了病叶粗汁液中的芜菁花叶病毒(TuMV)、大豆花叶病毒(sMV)和黄瓜花叶病毒(CMV),病叶粗汁液可被检测的最大稀释度分别为1:5120、1:2560和1:1280。提纯的大豆花叶病毒和黄瓜花叶病毒可检测的最低限量分别为1.7ng和1.2ng。以牛血清白蛋白、吐温和聚乙烯吡咯啉酮作封闭液,均可获得满意的结果。应用斑点法检测芜菁花叶病毒和大豆花叶病毒时,其抗血清稀释1:500倍可获得满意效果,稀释2000倍仍可用于检测。

Detection of Plant Viruses by Dot-immunobinding Assay

TuMV, SMV, and CMV were detected by dot-immunobinding assay (DIA) at dilutions of 1:5120, 1:2560 and 1:1280, respectively. DIA successfully detected 1.7ng of purified SMV and 1.2ng of purified CMV. DIA was used with horseradish peroxidase-linked goat anti-rabbit IgG and nitrocellulose membrane was used as the solid phase. 3, 3-Diamino-benzidine was used as the suhstrate and got good results. Absorption of the first antibody with healthy plant sap was necessary to eliminate background color. This assay is simple, rapid and sensitive and should prove to be a useful and practical diagnostic technique for plant virus diseases.