Abstract
The purpose of this study was to adapt an ELISA test for diagnosing of “Apple Chlorotic Leaf Spot Virus” (CLSV) in apple trees. This work was carried out at Centro de Pesquisa Agropecuária de Clima Temperado–CPACT/EMBRAPA, Pelotas–RS, Brazil, during the 1996 spring season. The application of ADGEN Diagnostic Systems protocol does not give some positive results from diseased apple trees. The procedure modified by FLEGG & CLARK (1979) gives an unsatisfactory result for color reaction in the positive samples. It means it is necessary to adapt this methodology. When the antigen was obtained from leaves grown from the base to the intermediate position in the stem and grounded with extracting buffer—0.02 M, pH 7.4 (1 g tissue: 3 ml extracting buffer) and polyclonal antisera and antibody alkaline phosphatase conjugate was diluted in coating buffer—0.05 M, pH 9.6 (1 μg antisera or antibody: 500 μl coating buffer) the reaction become more intensive and the test was able to diagnosticate the presence of the pathogen in infected leaves of apple trees.
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