Abstract
Production and diagnostic utility of monoclonal antibody (MCA) to zucchini yellow mosaic virus (ZYMV) in enzyme-linked immunosorbent assay (ELISA) are described. Four selected hybridomas produced MCA of the IgG2b subclass, and the titre of these MCA in ascitic fluid ranged between 107 and 109 which were 1, 000 times higher than those in culture fluid. Of the six procedures of ELISA tested, the most sensitive result was obtained when the plates were coated with MCA and rabbit polyclonal antibody (PCA) was used as intermediate antibody (procedure 5). This assay was able to detect ZYMV in purified preparation at detection end point of 0.11ng/ml and in crude extracts of pumpkin leaves at dilution end point of 106107. The use of MCA ZYMV-45 in non-precoated I-ELISA could discriminate ZYMV and watermelon mosaic virus 2 (WMV-2). Thus, the result represented a major advance in the detection of ZYMV which the use of PCAs in the same assay could not discriminate these two viruses. This assay gave also strong positive reactions with all of ZYMV-infected samples that reacted with PCA in double-antibody sandwich ELISA, thereby providing specific diagnostic tool for ZYMV which is needed for guiding the management of potyviruses on cucurbits.
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