斑点免疫结合法检测3种植物病毒

研究了改进的斑点免疫结合法在检测芜菁花叶病毒、烟草花叶病毒和小西葫芦黄化花叶病毒中的应用.无论是使用全抗血清还是纯化的免疫球蛋白,检测感染组织粗汁液中的病毒或纯化的病毒,均获得了满意的结果.同时进行的斑点免疫结合试验、酶联免疫吸附试验和免疫吸附电子显微术等的比较结果表明:无论是检测纯化病毒还是感染组织粗汁液中的病毒,斑点免疫结合法的检测敏感性均优于后2种方法,病毒的可测感度芜菁花叶病毒为0.65ng,烟草花叶病毒为0.39 ng,小西葫芦黄化花叶病毒为0.45 ng;应用碱性磷酸酶标记抗体及羟基吲哚磷酸盐和氮兰四唑为底物较为合适,这种底物可在室温下长期保存,并且反应产物为不褪色的紫色,易于观察和保存.

DETECTION OF THREE PLANT VIRUSES USING DOT-IMMUNOBINDING ASSAY

The detection of Turnip mosaic virus, Tobacco mosaic virus and Zucchini yellow mosaic virus using modified dot-immunobinding assay is carried out. The results of the experiment are satisfactory regardless of whether crude virus infected extracts and purified virus preparation and whether crude antiserum or purified IgG is used. The sensitivity of enzyme-linked immunosorbent assay (ELISA), immunosorbent electron microscopy (ISEM) and dot-immunobinding assay (DIBA) are compared with purified viruses and crude extracts of TuMV, TMV and ZYMV. DIBA assay is more sensitive than ELISA and ISEM, DIBA can detect as little as 0.65 ng of purified TuMV, 0.39 ng of purified TMV, 0.45 ng of purified ZYMV and is superior to other two methods. The combination of IgG-conjugated alkaline phosphatase and indoxyl phosphate and nitroblue tetrazolium as the substrates gives very good color reaction, wich can be distinguished by naked eyes without false positive.