A biotinylated DNA probe to detect bacterial cells in artificially contaminated foodstuffs.
A biotin-labelled DNA probe was used in a dot-blot hybridization test to demonstrate the presence of Escherichia coli in a variety of artificially contaminated foodstuffs. Positive hybridization was detected by using a streptavidine/polyalkaline phosphatase conjugate to generate an insoluble coloured precipitate in the presence of an appropriate dye. The colour intensity was measured with a computer-controlled image analysis system which assessed objectively the hybridization signal produced by each sample. The method was capable of distinguishing positive hybridization at cell concentrations exceeding 10(4) cells/dot-blot, equivalent to 2 x 10(7) cells/g food, and had none of the drawbacks normally associated with the use of radioactively labelled DNA in hybridization techniques. The procedure is highly specific and takes less than 30 h. Many samples can be screened simultaneously and the procedure can be used to detect any species for which a suitable DNA probe is available.
Dovey
S
S
Department of Microbiology, University Hospital, Nottingham, UK.
Towner
K J
KJ
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D001710:Biotin; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis*; D005516:Food Microbiology; D007091:Image Processing, Computer-Assisted; D009693:Nucleic Acid Hybridization; D011237:Predictive Value of Tests
D015342:DNA Probes; D004269:DNA, Bacterial; D001710:Biotin