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Automated pneumococcal MLST using liquid-handling robotics and a capillary DNA sequencer

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Abstract

Multilocus sequence typing (MLST) is used by the Scottish Meningococcus and Pneumococcus Reference Laboratory (SMPRL) as a routine method for the characterization of certain bacterial pathogens. The SMPRL recently started performing MLST on strains of Streptococcus pneumoniae, and here we describe a fully automated method for MLST using a 96-well-format liquid-handling robot and a 96-capillary automated DNA sequencer.

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References

  1. Maiden, M.C., Bygraves, J. A., Feil, E., et al. (1998) Multilocus sequence typing: A portable approach to the identification of clones within populations of pathogenic microorganisms. Proc. Natl. Acad. Sci. USA 95, 3140–3145.

    Article  PubMed  CAS  Google Scholar 

  2. Enright, M.C. and B.G. Spratt (1998) A multilocus sequence typing scheme for Streptococcus pneumoniae: Identification of clones associated with serious invasive disease. Microbiology 144(Pt 11), 3049–3060.

    Article  PubMed  CAS  Google Scholar 

  3. Clarke, S.C., M.A. Diggle, and G.F. Edwards (2001) Semiautomation of multilocus sequence typing for the characterisation of clinical isolates of Neisseria meningitidis. J. Clin. Microbiol. 39, 3066–3071.

    Article  PubMed  CAS  Google Scholar 

  4. Smart, L.E. (1986) Serotyping of Streptococcus pneumoniae strains by coagglutination. J. Clin. Pathol. 39, 328–331.

    Article  PubMed  CAS  Google Scholar 

  5. Smart, L.E. and J. Henrichsen (1986) An alternative approach to typing of Streptococcus pneumoniae strains by coagglutination. Acta. Pathol. Microbiol. Immunol. Scand. [B]. 94, 409–413.

    CAS  Google Scholar 

  6. Clarke, S.C. and M.A. Diggle (2002) Automated PCR/sequence template purification. Mol. Biotechnol. 21, 221–224.

    Article  PubMed  CAS  Google Scholar 

  7. Diggle, M.A. and S.C. Clarke (2002) Rapid assignment of nucleotide sequence data to allele types for multi-locus sequence analysis (MLSA) of bacteria using an adapted database and modified alignment program. J. Mol. Microbiol. Biotechnol. 4, 515–517.

    PubMed  CAS  Google Scholar 

  8. Clarke, S.C., Denham, B.C., Reid, J.A., et al. (2002) Pneumococcal disease in Scotland 1999. SCIEH Weekly Rep. 36, 62–63.

    Google Scholar 

  9. Kalin, M. (1998) Pneumococcal serotypes and their clinical relevance. Thorax 53, 159–162.

    Article  PubMed  CAS  Google Scholar 

  10. Scott, J.A., et al. (1996) Serogroup-specific epidemiology of Streptococcus pneumoniae: Associations with age, sex, and geography in 7,000 episodes of invasive disease. Clin. Infect. Dis. 22, 973–981.

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Microbiology, Stobhill Hospital, Scottish Meningococcus and Pneumococcus Reference Laboratory, North Glasgow University Hospital NHS Trust, Balornock Rd., G21 3UW, Glasgow, UK

    Johanna Jefferies, Stuart C. Clarke & Tim Mitchell

  2. Institute of Biomedical and Life Sciences, University of Glasgow, UK

    Stuart C. Clarke & Mathew A. Diggle

  3. Glasgow Dental School and Hospital, Glasgow

    Andrew Smith

  4. Department of Biological Sciences, Warwick University, UK

    Chris Dowson

Authors
  1. Johanna Jefferies
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  2. Stuart C. Clarke
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  3. Mathew A. Diggle
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  4. Andrew Smith
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  5. Chris Dowson
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  6. Tim Mitchell
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Corresponding author

Correspondence to Stuart C. Clarke.

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Jefferies, J., Clarke, S.C., Diggle, M.A. et al. Automated pneumococcal MLST using liquid-handling robotics and a capillary DNA sequencer. Mol Biotechnol 24, 303–307 (2003). https://doi.org/10.1385/MB:24:3:303

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  • DOI: https://doi.org/10.1385/MB:24:3:303

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