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

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

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    12 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)335-341
    Number of pages7
    JournalJournal of Antimicrobial Chemotherapy
    Volume20
    Issue number3
    DOIs
    Publication statusPublished - Sep 1987

    Bibliographical note

    Funding Information:
    This work was supported by a research grant from the Trent Regional Health Authority.

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