Optimization of a Chemiluminescent Dot-Blot Immunoassay for Detection of Potato Viruses |
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| Authors: | Ana C Fulladolsa Palma Rajitha Kota Amy O Charkowski |
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| Institution: | 1. Department of Plant Pathology, University of Wisconsin-Madison, 1630 Linden Dr., Madison, WI, 53706, USA
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| Abstract: | Vegetative propagation of potato leads to virus accumulation, resulting in significant yield losses and reduced quality. Virus identification is critical for developing disease management strategies and measuring seed lot health. The most widely used method of virus diagnosis in seed potatoes is a post-harvest test, for which the enzyme-linked immunosorbent assay (ELISA) is often used. ELISA was previously modified by substituting microtiter plates with membranes to develop a more flexible and inexpensive assay. We optimized a dot-blot immunoassay with viral proteins bound to a polyvinylidene fluoride (PVDF) membrane and detection of the proteins with alkaline phosphatase-labeled antibodies and a chemiluminescence reagent. The assay was tested for detection of viruses of seven genera. We have also altered the assay by spotting an antibody array onto a PVDF membrane and tested it for its potential uses as a diagnostic tool for Potato virus Y, Hosta virus X, and Potato leafroll virus. |
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