[Molecular biological method cascade for detection, specific isolation and characterization of shiga toxin-producing Escherichia coli (STEC) in food].
Shigatoxin-producing E. coli (STEC) and their subgroup, the enterohaemorrhagic E. coli (EHEC) are known since about 25 years. Only EHEC can cause diseases in humans. A molecularbiological method cascade is described in this article. It contains the following steps: preenrichment, enrichment, preparation of samples (DNA extraction), screening PCR, specific STEC isolation by using DNA DNA hybridization, verification of isolates as STEC by using PCR, characterization of STEC isolates. Pulsed field gel electrophoresis and DNA sequencing are additional methods for the detection of clonal correlations. They should be performed in special labs, only. Furthermore, some information about an interlaboratory ring trial within Germany and some conclusions are given.
Gallien
Peter
P
Bundesinstitut für Risikobewertung (BfR), Nationales Veterinärmedizinisches Referenzlabor für E. coli, Dessau.
D000818:Animals; D001483:Base Sequence; D015169:Colony Count, Microbial; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D016521:Electrophoresis, Gel, Pulsed-Field; D016021:Epidemiologic Studies; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004927:Escherichia coli Infections / Q000175:diagnosis* / Q000382:microbiology; D019453:Escherichia coli O157 / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D005506:Food Contamination / Q000032:analysis; D005516:Food Microbiology; D006801:Humans; D016133:Polymerase Chain Reaction; D022603:Shiga Toxins / Q000032:analysis* / Q000096:biosynthesis
D015342:DNA Probes; D004269:DNA, Bacterial; D022603:Shiga Toxins