ISHS Acta Horticulturae 217: IV International Workshop on Fire Blight
DIRECT COUNTS OF ERWINIA AMYLOVORA BY IMMUNOFLUORESCENT STAINING WITH MONOCLONAL ANTIBODIES |
| Authors: | C.P. Lin, T.A. Chen, J.M. Wells, T. van der Zwet |
| Abstract:
A modified direct-count technique was applied in the identification and detection of E. amylovora by immunofluorescent staining with monoclonal antibodies (MA). Bacterial suspension prepared from pure cultures or from rinsed infected pome fruit blossoms were pelleted in a microcentrifuge tube at 13,000 g for five min. Bacterial pellets were resuspended and incubated in MA for one hour, then centrifuged and resuspended in Fluorescein isothiocyanate (FITC) conjugated antimouse immunoglobulin for another one hour. Bacteria were then collected on polycarbonate membranes by filtration and examined under an epi-fluorescent microscope. This technique can conveniently detect 5 x 103 cells and is useful in detecting and monitoring the infection of E. amylovora in pome fruit trees and fruits. | |
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