Identification of Shiga-like toxin type II producing Escherichia coli using the polymerase chain reaction and a digoxigenin labelled DNA probe

Mol Cell Probes. 1992 Jun;6(3):209-14. doi: 10.1016/0890-8508(92)90018-s.

Abstract

Epidemiological studies have demonstrated that enterohaemorrhagic strains of Escherichia coli which cause the haemolytic uremic syndrome in humans and the oedema disease in pigs more frequently produce Shiga-like toxin type II (SLT-II) than any other member of the Shiga-like toxin family. A technique has been developed for the identification of SLT-II producing E. coli using the polymerase chain reaction (PCR) and a digoxigenin (DIG)-labelled DNA probe to facilitate the early detection and epidemiological analysis of these pathogens. Whole cell DNA liberated from isolated colonies during the denaturation step of PCR was amplified using a primer pair which is homologous to the slt-II gene sequences. The amplification products were transferred directly to a nitrocellulose membrane or following agarose gel electrophoresis and DNA denaturation. A chemically labelled DNA probe, prepared using PCR with the incorporation of DIG, was used to identify the PCR products of strains which produced SLT-II or a variant of SLT-II.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Toxins / analysis*
  • Bacterial Toxins / genetics
  • Blotting, Southern
  • DNA Probes*
  • Digoxigenin*
  • Enterotoxins / analysis*
  • Enterotoxins / genetics
  • Escherichia coli / chemistry*
  • Escherichia coli / genetics
  • Escherichia coli / isolation & purification
  • Escherichia coli Infections / diagnosis
  • Escherichia coli Infections / microbiology
  • Genes, Bacterial
  • Humans
  • Nucleic Acid Hybridization
  • Polymerase Chain Reaction*
  • Sensitivity and Specificity
  • Shiga Toxin 2

Substances

  • Bacterial Toxins
  • DNA Probes
  • Enterotoxins
  • Shiga Toxin 2
  • Digoxigenin