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[Utilization of nylon membranes for specific isolation and characterization of verotoxin-producing Escherichia coli
P Gallien1, H Klie, K W Perlberg
1Bundesinstitut für gesundheitlichen Verbraucherschutz und Veterinärmedizin, Standorte in Dessau.
Abstract:
A method for specific isolation of VT(+)-strains in raw milk is given. DNA-hybridization technique with DIG-labeled PCR-amplificates as probes are the basis. No background is seen by using "DIG Easy Hyb" solution and nylon membranes for colony- and plaque-hybridization (Boehringer Mannheim GmbH). Marked colonies are visible on the membranes after detection. So it is possible to select these colonies from a masterplate. The results are available within one day (without enrichment and membrane preparation). After stripping the membranes can be used for a new hybridisation to detect another factor of virulence.