Abstract
A simple kit for rapid detection of potato virus Y by latex serological test was developed. The test is carried out on a white cardboard sheet and the results can be read by naked eye in two minutes. A test card of 10 x 6 cm holds latex sensitized antibodies, buffers and other necessary ingredients as dry blue colored formulate on the ringed areas of the card. A test card includes space for six tests and positive and negative controls. The kit also includes disposable plastic sticks for mixing the samples with test reagents and a hand press with disposable plastic tips. For testing, dried reagents are dissolved in drops of sample and mixed. After gentle rotation, samples containing virus appear clearly granulated while samples from healthy plants remain unagglutinated. The testing of undiluted extracts of evenly developed tuber sprouts resulted in over 91 % identity with the results obtained with ELISA that was used as a control method. Testing of diluted leaf extracts reached the same reliability but undiluted leaf extracts from glasshouse grown potatoes were not well suitable as test samples because of their dark green color. No such problems occurred with field grown material and a complete identity with the ELISA readings was true when the samples included secondarily infected potato plants. No reaction to other potato viruses than PVY was obtained by the test kit.
Highlights
Serological diagnosis of plant viruses has developed drastically during the past decade, with the introduction of labelled antibody techniques, especially the ELISA
The ELISA has a number of advantageous properties over the methods previously used for routine detection of plant viruses, above all high sensitivity and specificity, and it is easy to be applied in any moderately equipped research laboratory
Immunoglobulins from our high-titre antisera were effective in coating latex particles at concentrations as low as 10—20/tg/ml
Summary
Serological diagnosis of plant viruses has developed drastically during the past decade, with the introduction of labelled antibody techniques, especially the ELISA (enzyme-linked immunosorbent assay, Clark and Adams 1977). The ELISA has a number of advantageous properties over the methods previously used for routine detection of plant viruses, above all high sensitivity and specificity, and it is easy to be applied in any moderately equipped research laboratory. It cannot be used for field diagnosis, and it takes one to one and a half days to have the results. The ELISA is the only relatively rapid test method for the detection of potato virus Y (PVY). Other more traditional techniques such as the agglutination test, microprecipitin test (van Slogteren 1957), and radial immunodiffusion test (Shepard 1972, Richter et al 1979 a), are not sensitive enough for the routine detection of PVY in potato tubers or leaves
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