[Utilization of nylon membranes for specific isolation and characterization of verotoxin-producing Escherichia coli using DNA probes].
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.
Gallien
P
P
Bundesinstitut für gesundheitlichen Verbraucherschutz und Veterinärmedizin, Standorte in Dessau.
Klie
H
H
Perlberg
K W
KW
Protz
D
D
D000818:Animals; D001427:Bacterial Toxins / Q000096:biosynthesis*; D002417:Cattle; D015342:DNA Probes; D004768:Enterotoxins / Q000096:biosynthesis; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D008567:Membranes, Artificial; D008892:Milk / Q000382:microbiology*; D009757:Nylons; D016133:Polymerase Chain Reaction / Q000379:methods; D022622:Shiga Toxin 1
D001427:Bacterial Toxins; D015342:DNA Probes; D004768:Enterotoxins; D008567:Membranes, Artificial; D009757:Nylons; D022622:Shiga Toxin 1