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Fimbrial-Specific Monoclonal Antibody-Based ELISA for European Potato Strains of Erwinia chrysanthemi and Comparison to PCR

A murine hybridoma cell line, named 6A6, was developed to produce monoclonal antibodies for serological detection of European potato strains of Erwinia chrysanthemi. The monoclonal antibodies were of the immunoglobulin G_2b type and were shown to react with a fimbrial antigen by immuno-gold electron microscopy, and with the fibrillin protein by Western blotting. In enzyme-linked immunosorbent assay (ELISA), the monoclonal antibody reacted with all but two strains of E. chrysanthemi isolated from potato. One non-reactive strain originated from Australia and therefore was likely a different biovar, and the other strain was of unknown origin. The monoclonal antibody also reacted with 20 out of 36 strains of E. chrysanthemi isolated from hosts other than potato. A triple-antibody ELISA test utilizing monoclonal antibody 6A6 successfully detected E. chrysanthemi in infected potato stems and tubers but sensitivity was limited to about 10⁷ CFU/ml, compared to a sensitivity of 10³ CFU/ml for a polymerase chain reaction test using published primers directed to the pectate lyase gene.