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Multilocus characterization scheme for shiga toxin-encoding bacteriophages.

Darren L Smith1, Brian M Wareing, Paul C M Fogg

  • 1School of Biological Sciences, Microbiology Research Group, University of Liverpool, Liverpool L69 7ZB, United Kingdom. hallison@liverpool.ac.uk.

Applied and Environmental Microbiology
|October 24, 2007
PubMed
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A new PCR method types Shiga toxin-encoding bacteriophages (Stx phages) found in foodborne Shiga toxin-producing Escherichia coli (STEC). This tool helps understand Stx phage diversity and their role in STEC epidemiology.

Area of Science:

  • Microbiology
  • Virology
  • Food Safety

Background:

  • Shiga toxin-producing Escherichia coli (STEC) are significant foodborne pathogens.
  • Shiga toxin (Stx) production is mediated by integrated lambdoid bacteriophages (Stx phages).
  • Stx phages exhibit considerable genetic and morphological diversity.

Purpose of the Study:

  • To design and validate a PCR-based multilocus typing scheme for Stx phages.
  • To characterize the genetic diversity of Stx phages within STEC isolates.
  • To enable robust epidemiological studies of Stx phages.

Main Methods:

  • Development of PCR primer sets targeting key lambdoid bacteriophage genes.
  • Application of the typing scheme to control phages and 70 Stx phage preparations from STEC isolates.

Related Experiment Videos

  • Analysis of genetic diversity and gene heterogeneity within Stx phage populations.

    • Main Results:

    • The PCR scheme successfully profiled purified Stx phages and identified lambdoid phage-borne genes.
    • Significant genetic diversity was observed among Stx phages.
    • A predominance of short-tailed phages with a conserved tail spike protein gene was noted.

    Conclusions:

    • The developed multilocus typing scheme is effective for characterizing Stx phages.
    • This method aids in identifying Stx phages in both induced and prophage forms within STEC.
    • The findings provide a foundation for future epidemiological investigations into the impact of Stx phages on STEC.