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Journal of Antimicrobial Chemotherapy (1987) 20, 335-341
© 1987 The British Society for Antimicrobial Chemotherapy

research-article

Rapid detection of a specific trimethoprim resistance gene using a biotinylated DNA probe

G. I. Carter, K. J. Towner* and R. C. B. Slack

Department of Microbiology and PHLS Laboratory, University Hospital, Queen's Medical Centre Nottingham NG7 2UH, UK

Received 9 March 1987;

*Corresponding author

A DNA probe specific for the dihydrofolate reductase (DHFR) type I gene was labelled with biotin by the process of nick-translation and used to screen 83 independently-derived trimethoprim R plasmids from Enterobacteriaceae. Hybridization was detected using streptavidine and a biotin-conjugated alkaline phosphatase to generate an insoluble coloured precipitate following the addition of an appropriate dye. Sixty-eight plasmids (81.9%) hybridized with the probe for DHFR type I. The method could be adapted for use with any antibiotic resistance gene for which a suitable DNA probe is available and has none of the drawbacks associated with the use of radioactively-labelled DNA in hybridization techniques.


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