Development of an Immunosorbant Assay for Seedborne Clavibacter michiganensis subsp. nebraskense in Corn Seeds

Abstract:

We purified the McAbs and then determined the subclasses, activity and specificity of the Clavibacter michiganensis subsp. nebraskense McAbs (4H4 and 4G12). We compared the detection sensitivity between indirect ELISA and double antibody sandwich ELISA (DAS-ELISA) and utilized both of them on the detection of the corn bacteria. Titers of both McAbs in 0.4 g/L were 1 : 256000, subclasses of 4H4 belongs to IgG2a while 4G12 belongs to IgG2b. No cross reactivity was found within 16 bacterial strains tested from corn and other plants by ELISA. The minimun detection limit of the bacteria in the seed suspension with DAS-ELISA was 1.0 × 109 CFU/L. A specific and sensitive DAS-ELISA for detection of the C. michiganensis subsp. nebraskense was developed.

Key words:C. michiganensis subsp. nebraskense, monoclonal antibody, ELISA