Study on detection of Pantoea stwartii subsp. stewartii based on Dot-ELISA

Abstract:

Enzyme linked immunosorbent assay performing on newly modified nitrocellulose membrane strip (NCM test strip) for detection of Pantoea stwartii subsp.stewartii, the casual agent of bacterial wilt of corn, was established. The test strip used in this study has eight round pieces of nitrocellulose membrane with a diameter of 2 mm, which was cut with a VersaLaser, and then was affixed on a plastic adhesive tape. Detection sensitivity for the pathogen was increased by 10-100 folds when 10μL per site of the bacterial suspension was used, as compared to that in the normal amount of 1uL per dot; detection sensitivity in indirect dot-ELISA was 10 folds higher than that in sandwich double antibodies dot-ELISA (DAS-dot- ELISA)，These results were further confirmed from the ELISA tests carried on the normal microtiter plates. This study will promote development of microfluidic chip detection method combined with the dot immunoassay for P. stwartii subsp.stewartii on corn.