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Potato is a host for many viruses and vegetative propagation of the crop leads to their accumulation, resulting in significant yield losses and reduced quality. The most widely used method of diagnosis of viral infections is the post-harvest test, for which the enzyme-linked immunosorbent assay (ELISA) is used. This method is efficient, and relatively inexpensive, but the large number of samples processed means that even small improvements can result in significant savings. Others have modified the ELISA by substituting microtiter plates for polyvinylidene fluoride (PVDF) membranes to develop a more flexible and inexpensive assay. We optimized a dot-blot immunoassay with viral proteins bound to a 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. The cost of this assay is 85% less than that of a standard ELISA. 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 plant viruses. We used the dot-blot immunoassay for detection of <i>Potato virus Y</i> (PVY) in 1,530 samples from a post-harvest test to determine virus incidence in tubers after a field trial. Of the 110 PVY-positive samples, 101 were detected by using the assay, while the remaining 9 were not detectable due to high background signal or blotching, and were identified by using ELISA. <p><p>Keywords: Virus-Viroid, Root-Tuber Crops, Potato