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Journal of Clinical Microbiology, October 1999, p. 3402-3404, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Elimination of Bacterial DNA from Taq DNA Polymerases by Restriction Endonuclease Digestion

Nora M. Carroll,dagger Peter Adamson, and Narciss Okhravi*

Department of Clinical Ophthalmology, The Institute of Ophthalmology, London EC1V 9EL, United Kingdom

Received 2 April 1999/Returned for modification 4 June 1999/Accepted 6 July 1999

The incidence of false positives due to the presence of bacterial DNA in Taq DNA polymerase is an obstacle to the use of PCR in the diagnosis of infection. We describe a method that uses a restriction enzyme to destroy the ability of contaminating sequences to act as templates for a nested PCR which uses primers based on the 16S rRNA genes. The method was used prior to a PCR that amplified 10 fg of bacterial DNA. This method can be readily adapted to suit other sensitive PCRs required for clinical applications.

* Corresponding author. Mailing address: Department of Clinical Ophthalmology, The Institute of Ophthalmology, Bath St., London EC1V 9EL, United Kingdom. Phone: 0171-608-6872. Fax: 0171-608-6931. E-mail: nokhravi{at}menu.hgmp.mrc.ac.uk.

dagger Present address: Department of Medical Biochemistry, University of Stellenbosch, Tygerberg 7505, South Africa.

Journal of Clinical Microbiology, October 1999, p. 3402-3404, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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