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Nucleotide sequence-based typing of meningococci directly from clinical samples
Mathew A Diggle1, Carolyn M Bell1, Stuart C Clarke1
1Scottish Meningococcus and Pneumococcus Reference Laboratory, Department of Microbiology, House on the Hill, Stobhill Hospital, Balornock Road, Glasgow G21 3UW, UK 2Faculty of Biomedical and Life Sciences, University of Glasgow, UK#dReceived 19 September 2002 Accepted 25 February 2003.
Journal of Medical Microbiology
|May 16, 2003
Summary
This study introduces a new method for identifying meningococcal disease (MD) using multilocus sequence typing (MLST) directly from patient samples. This advance improves the surveillance of Neisseria meningitidis, even without a culture isolate.
Area of Science:
- Microbiology
- Epidemiology
- Molecular Biology
Background:
- Meningococcal disease (MD) outbreaks are challenging to monitor due to unpredictable characteristics, causing public anxiety.
- Traditional molecular techniques for MD epidemiology are often specialized, labor-intensive, and lack reproducibility.
- Multilocus sequence typing (MLST) offers an improvement by utilizing unambiguous nucleotide sequencing data for microbial characterization.
Purpose of the Study:
- To describe the application of MLST for laboratory confirmation and characterization of Neisseria meningitidis directly from clinical samples.
- To address the need for enhanced MD surveillance following the introduction of serogroup C conjugate vaccines.
- To enable typing of the infecting organism in the absence of a culture isolate.
Main Methods:
- Development and application of a newly designed set of primers for MLST.
- Primers are complementary to nucleotide sequences external to existing culture-based MLST primers for meningococci.
- Utilized MLST for genotypic characterization directly from clinical samples.
Main Results:
- A highly sensitive procedure was developed for efficient genotypic characterization of meningococci.
- The method allows for direct characterization from clinical samples, bypassing the need for culture isolates.
- Successful application of MLST for laboratory confirmation and characterization of Neisseria meningitidis.
Conclusions:
- The described MLST approach provides a sensitive and efficient method for characterizing Neisseria meningitidis directly from clinical samples.
- This technique enhances the surveillance capabilities for meningococcal disease, particularly in the post-vaccine era.
- The method overcomes limitations of traditional techniques, offering improved reproducibility and reduced labor intensity.