Articles

DETECTION OF APPLE CHLOROTIC LEAF SPOT VIRUS AND APPLE STEM GROOVING VIRUS BY DOT-IMMUNOBINDING ASSAY

Article number
472_2
Pages
51 – 54
Language
Abstract
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 μl of horseradish peroxidase scale protein A(HRP-SPA) and 25 μl 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.

Publication
Authors
Q. Wang, Y. Shi, J. Yang, Q. Sun
Keywords
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