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Automated pneumococcal MLST using liquid-handling robotics and a capillary DNA sequencer
Johanna Jefferies1, Stuart C Clarke, Mathew A Diggle
1Scottish Meningococcus and Pneumococcus Reference Laboratory, North Glasgow University Hospital NHS Trust, Department of Microbiology, Stobhill Hospital, Balornock Rd., Glasgow, G21 3UW, UK.
Molecular Biotechnology
|June 5, 2003
Summary
The Scottish Meningococcus and Pneumococcus Reference Laboratory (SMPRL) developed an automated method for bacterial characterization. This approach uses robotics and DNA sequencing for Multilocus Sequence Typing (MLST) of Streptococcus pneumoniae.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Multilocus Sequence Typing (MLST) is a standard method for bacterial pathogen characterization.
- The Scottish Meningococcus and Pneumococcus Reference Laboratory (SMPRL) routinely employs MLST.
- There is a need for efficient and high-throughput methods in bacterial typing.
Purpose of the Study:
- To describe a fully automated method for MLST.
- To adapt MLST for high-throughput analysis of Streptococcus pneumoniae strains.
- To integrate liquid-handling robotics and automated DNA sequencing for bacterial characterization.
Main Methods:
- Utilized a 96-well-format liquid-handling robot for automated sample processing.
- Employed a 96-capillary automated DNA sequencer for high-throughput genetic analysis.
- Applied MLST to characterize strains of Streptococcus pneumoniae.
Main Results:
- Successfully implemented a fully automated MLST workflow.
- Demonstrated the feasibility of high-throughput bacterial typing using robotic automation.
- Generated genetic profiles for Streptococcus pneumoniae strains efficiently.
Conclusions:
- The described automated method enhances the efficiency of MLST for bacterial characterization.
- Robotic automation and automated sequencing are suitable for routine high-throughput MLST.
- This approach facilitates large-scale epidemiological studies of bacterial pathogens like Streptococcus pneumoniae.