Rapid genotyping of Escherichia coli O157 isolates by random amplification of polymorphic DNA.

Abstract

Although several typing methods have been described for Shiga-like toxin-producing Escherichia coli O157, the methods are somewhat cumbersome. Using 22 isolates of Escherichia coli O157 and five other Escherichia coli isolates, two primers (M13 core sequence and 970-11) were found to give excellent differentiation between isolates using random amplified polymorphic DNA (RAPD). Using only the presence or absence of variable bands, a matrix of 20 variable characters was identified. From these characters, similarity coefficients were calculated and a phenogram constructed. All of the Escherichia coli O157 isolates were easily distinguished from the non-O157 Escherichia coli isolates. Using a 95% similarity cutoff, we found 13 RAPD types among the 22 Escherichia coli O157 isolates. Isolates thought to be identical by toxin and phage typing as well as by epidemiological association were distinguished, and others thought to be distinct by lack of epidemiological association were identical. RAPD using M13 and 970-11 primers is a potentially useful typing tool for Escherichia coli isolates of serotype O157 and possibly other Escherichia coli isolates.