Nucleotide sequence-based typing of meningococci directly from clinical samples.
The unpredictable characteristics of meningococcal disease (MD) make outbreaks complicated to monitor and consequently lead to high levels of public anxiety. Traditional molecular techniques have been utilized in order to understand better the epidemiology of MD, but some have disadvantages such as being highly specialized and labour-intensive, with low reproducibility. Some of these problems have been overcome by using multilocus sequence typing (MLST). This technique exploits the unambiguous nature and electronic portability of nucleotide sequencing data for the characterization of micro-organisms. The need for enhanced surveillance of MD after the introduction of serogroup C conjugate vaccines means that it is important to gain typing information from the infecting organism in the absence of a culture isolate. Here, the application of MLST for the laboratory confirmation and characterization of Neisseria meningitidis directly from clinical samples is described. This involved using a newly designed set of primers that were complementary to nucleotide sequences external to the existing MLST primers already in use for culture-based MLST of meningococci. This combination has produced a highly sensitive procedure to allow the efficient genotypic characterization of meningococci directly from clinical samples.
Scottish 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.
Mathew A
MA
Diggle
Scottish 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.
Carolyn M
CM
Bell
Scottish 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.
Stuart C
SC
Clarke
D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry; D004196:Disease Outbreaks; D006801:Humans; D008589:Meningococcal Infections / Q000453:epidemiology / Q000382:microbiology*; D009345:Neisseria meningitidis / Q000145:classification* / Q000235:genetics; D016133:Polymerase Chain Reaction; D012606:Scotland / Q000453:epidemiology; D017422:Sequence Analysis, DNA / Q000379:methods*
D017931:DNA Primers; D004269:DNA, Bacterial