Development of a 5'-nuclease PCR assay for detecting Shiga toxin-producing Escherichia coli O145 based on the identification of an 'O-island 29' homologue

J Appl Microbiol. 2003;94(4):587-94. doi: 10.1046/j.1365-2672.2003.01872.x.

Abstract

Aims: A DNA sequence, from Escherichia coli STEC O145, homologous to O-island 29 from STEC O157 is described, together with a real-time PCR assay for detecting it.

Methods and results: PCR and sequencing were used to identify the 'O-island 29' homologous DNA sequence from STEC O145 (strain VTH34). The sequence divergence between the STEC O145 and O157 'O-island 29' allowed a STEC O145 5'-nuclease PCR assay to be developed.

Conclusions: The characterization of a novel locus in STEC O145 has allowed a specific O145 serogroup 5'-nuclease PCR assay to be designed.

Significance and impact of the study: These findings increase the number of serogroup PCR assays available as alternatives to classical O-serotyping of E. coli.

Publication types

  • Evaluation Study

MeSH terms

  • Amino Acid Sequence
  • Bacterial Typing Techniques / methods*
  • Base Sequence
  • DNA, Bacterial / genetics
  • Escherichia coli O157 / classification*
  • Escherichia coli O157 / metabolism
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Sequence Alignment
  • Shiga Toxin / biosynthesis*
  • Shiga Toxin / genetics

Substances

  • DNA, Bacterial
  • Shiga Toxin