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A biotinylated DNA probe to detect bacterial cells in artificially contaminated foodstuffs
Author(s) -
Dovey S.,
Towner K. J.
Publication year - 1989
Publication title -
journal of applied bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 0021-8847
DOI - 10.1111/j.1365-2672.1989.tb02452.x
Subject(s) - biotinylation , dot blot , dna , dna–dna hybridization , in situ hybridization , hybridization probe , conjugate , microbiology and biotechnology , contamination , southern blot , escherichia coli , chemistry , nucleic acid thermodynamics , biology , chromatography , biochemistry , gene , gene expression , base sequence , mathematics , mathematical analysis , ecology
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 2×10 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