Direct detection by PCR of Escherichia coli O157 and enteropathogens in patients with bloody diarrhea

Microbiol Immunol. 1997;41(10):819-22. doi: 10.1111/j.1348-0421.1997.tb01934.x.

Abstract

Direct detection of Escherichia coli O157 and foodborne pathogens associated with bloody diarrhea were achieved using polymerase chain reaction (PCR) after the preparation of DNA from stool specimens using the microspin technique. PCR was compared with cultivation and toxin production tests with respect to the efficiency of detection of each pathogen; E. coli O157, Vibrio parahaemolyticus, Salmonella serovar Enteritidis and Campylobacter jejuni. Detection of some or all of the above pathogens in clinical stool specimens was achieved using PCR. The minimum number of cells required for the detection of the above pathogens by PCR was 10(1) CFUs/0.5 g of stool sample. PCR was completed within 6 hr. The above pathogens were also detected in cultivation and toxin production tests. Partial purification of the template DNA using the microspin technique was essential for the elimination of PCR inhibitors from the DNA samples. This PCR method is an accurate, easy-to-read screening method for the detection of Shiga-like toxin producing E. coli O157 and enteropathogens associated with bloody diarrhea in stool specimens.

MeSH terms

  • Bacteria / genetics
  • Bacteria / isolation & purification*
  • Bacterial Infections / diagnosis
  • Bacterial Infections / microbiology
  • Campylobacter jejuni / isolation & purification
  • Colony Count, Microbial
  • DNA Primers
  • DNA, Bacterial / analysis
  • Diarrhea / diagnosis
  • Diarrhea / microbiology*
  • Electrophoresis, Agar Gel
  • Escherichia coli Infections / diagnosis
  • Escherichia coli Infections / microbiology
  • Escherichia coli O157 / isolation & purification*
  • Feces / microbiology*
  • Food Microbiology
  • Gastrointestinal Hemorrhage / microbiology
  • Genes, Bacterial / genetics
  • Humans
  • Polymerase Chain Reaction*
  • Salmonella enteritidis / isolation & purification
  • Vibrio parahaemolyticus / genetics
  • Vibrio parahaemolyticus / isolation & purification

Substances

  • DNA Primers
  • DNA, Bacterial