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Restriction endonuclease analysis of RAPD-PCR amplicons derived from Shiga-like toxin-producing Escherichia coli O157 isolates

Department of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT and *Life and Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET.

Corresponding author: Dr A.C. Hilton (e-mail: A.C.Hilton{at}aston.ac.uk).

Received 4 Feb. 2000; revised version received 28 May 2000; accepted 13 June 2000.

Abstract

Shiga-like toxin-producing Escherichia coli O157 isolates were characterised by random amplification of polymorphic DNA by PCR (RAPD-PCR) analysis developed to allow robust epidemiological typing of E. coli. Amplification with primer 1247 or 1290 generated a reproducible profile, but was not capable of distinguishing sufficiently between epidemiologically unrelated strains. Subsequent digestion of the amplicons with selected restriction endonucleases improved the discriminatory ability of this method for strains showing limited differentiation following RAPD-PCR analysis alone. Restriction endonuclease analysis of RAPD-PCR fragments generated from closely related strains has the potential to provide additional discriminatory information without loss of specificity.

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