Detection of PCR products of Escherichia coli O157:H7 in human stool samples using surface plasmon resonance (SPR)

FEMS Immunol Med Microbiol. 2000 Dec;29(4):283-8. doi: 10.1111/j.1574-695X.2000.tb01535.x.

Abstract

A method for the rapid detection of verotoxin-producing Escherichia coli O157:H7 in stools was evaluated. Strains possessing Shiga toxin-2 (stx-2) genes were isolated from stool samples and amplified using oligonucleotide primers. Stools spiked with cultured E. coli O157:H7 (strain 298 or strain 1646) were detected to be polymerase chain reaction (PCR) positive at 10(2) cfu per 0.1 g of stool. Stool samples from patients and healthy carriers showed a high correlation between positive results for a PCR and the presence of verotoxin-producing E. coli O157:H7, confirmed by isolation of serotype O157:H7 on sorbitol MacConkey medium (10 of 10 stool samples). These PCR products could be detected using a BIAcore 2000 surface plasmon resonance device using peptide nucleic acid as a sensor probe. In this report we use this method for the rapid detection of DNA from significant pathogenic organisms.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carrier State / microbiology
  • DNA, Bacterial / analysis
  • Escherichia coli Infections / microbiology
  • Escherichia coli O157 / genetics
  • Escherichia coli O157 / isolation & purification*
  • Feces / microbiology*
  • Humans
  • Peptide Nucleic Acids / genetics
  • Polymerase Chain Reaction / methods*
  • Shiga Toxin 2 / genetics*
  • Shiga Toxin 2 / metabolism
  • Surface Plasmon Resonance / methods*

Substances

  • DNA, Bacterial
  • Peptide Nucleic Acids
  • Shiga Toxin 2