Detection of apple chlorotic leaf spot virus and apple stem grooving virus by dot-immunobinding assay

文献类型： 外文期刊

第一作者： Wang, Q

作者： Wang, Q;Shi, Y;Yang, J;Sun, Q

作者机构：

期刊名称：17TH INTERNATIONAL SYMPOSIUM ON VIRUS AND VIRUS-LIKE DISEASES OF TEMPERATE FRUIT CROPS: FRUIT TREE DISEASES, VOLS 1 AND 2

ISSN： 0567-7572

年卷期： 1998 年 472 期

页码：

收录情况： SCI

摘要： Three thousand samples were collected from 72 cultivars, 10 stocks and 6 indicator plants infected with viruses and then apple chlorotic leafspot and apple stem grooving viruses were detected by dot-immunobinding assay (DIBA). Plant tissue was extracted with phosphate buffer saline (PBS), 0.05% Tween-20, 2% polyvinylpyrrolidone (PVP) and 0.2% bovine serum albumin (BSA) (w/v=1:2 for tree leaves, and w/v=1:1 for test tube seedlings). Equal amount of chloroform was added for centrifugation. Two microliters of crude extract and 25 mu l of horseradish peroxidase scale protein A(HRP-SPA) and 25 mu l of antibody (1/4000 to 1/6000 dilution) were used in DIBA. DIBA is simple, rapid (5 h), and consistent for a whole growing period. It is economical and convenient. The amount of antigen, antibody and HRP-SPA required for DIBA is only 1%, 2-3% and 12.5% of that for ELISA respectively. DIBA can be directly read without any instruments. Nitrocellulose membrane used for DIBA can be stored for later reading.

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