Restriction endonuclease analysis of RAPD-PCR amplicons derived from Shiga-like toxin-producing Escherichia coli O157 isolates.
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.
Hopkins
Katie L
KL
Hilton
Anthony C
AC
Department of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT and *Life and Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET.
D015373:Bacterial Typing Techniques / Q000379:methods; D017931:DNA Primers; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D019453:Escherichia coli O157 / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*; D022603:Shiga Toxins / Q000096:biosynthesis*
D017931:DNA Primers; D004269:DNA, Bacterial; D022603:Shiga Toxins; D004262:DNA Restriction Enzymes