Detection of enterotoxigenic Escherichia coli by DNA colony hybridization

Abstract

A method fo detecting large numbers of isolates of enterotoxigenic Escherichia coli is described in which the genes encoding th enterotoxins are detected, rather than the toxins themselves. Radiolabeled fragments of DNA encoding the heat-labile (LT) or heat-stable (ST) toxins were used as hybridization probes for homologous DNA sequences in E. coli colonies grown and lysed in situ on nitrocellulose filters. The LT probe detected all of 31 E. coli strains producing ST and LT or only LT, while the ST probe detected 12 of 17 strains producing only ST and three of 26 strains producing ST and LT. These results suggest that the LTs produced by different isolates of E. coli are homologous and that human isolates of E. coli produce at least two heterologous STs detectable in the infant mouse assay. The hybridization method also detected the presence of enterotoxigenic E. coli in bacterial growth in directly spotted stools from patients with acute diarrhea