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

G. I. Carter; K. J. Towner; R. C.B. Slack

doi:10.1093/jac/20.3.335

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 journal › Article › peer-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 language	English
Pages (from-to)	335-341
Number of pages	7
Journal	Journal of Antimicrobial Chemotherapy
Volume	20
Issue number	3
DOIs	https://doi.org/10.1093/jac/20.3.335
Publication status	Published - Sept 1987
Externally published	Yes

Bibliographical note

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

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title = "Rapid detection of a specific trimethoprim resistance gene using a biotinylated DNA probe",

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.",

author = "Carter, {G. I.} and Towner, {K. J.} and Slack, {R. C.B.}",

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

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AU - Towner, K. J.

AU - Slack, R. C.B.

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PY - 1987/9

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AB - 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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JF - Journal of Antimicrobial Chemotherapy

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

Abstract

Bibliographical note

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