Semiautomation of multilocus sequence typing for the characterization of clinical isolates of Neisseria meningitidis.
The Scottish Meningococcus and Pneumococcus Reference Laboratory (SMPRL) provides a national service for the laboratory confirmation of meningococcal and pneumococcal disease in Scotland. Part of this service includes the serogrouping of meningococcal isolates followed by typing and subtyping. The procedures for this are labor-intensive but important for the identification of linked cases and the surveillance of disease so that effective public health measures can be taken. However, different strains of meningococci, such as those within the electrophoretic type 37 complex, occurring during case clusters of disease are now indistinguishable by current methods. The SMPRL has started using multilocus sequence typing (MLST) as a routine method for the characterization of isolates of Neisseria meningitidis. MLST produces nucleotide sequence data of seven housekeeping genes providing results that are useful for public health management. However, the method is laborious and time-consuming and therefore lends itself towards automation. The SMPRL therefore developed a semiautomated method for MLST using a 96-well format liquid handler and an automated DNA sequencer. Semiautomated MLST is now provided as a reference service for Scotland. This work describes the methodology required for the characterization of N. meningitidis and highlights its usefulness for public health intervention.
Clarke
S C
SC
Scottish Meningococcus and Pneumococcus Reference Laboratory, North Glasgow University Hospital NHS Trust, Stobhill Hospital, Glasgow, United Kingdom. stuart.clarke@northglasgow.scot.nhs.uk
Diggle
M A
MA
Edwards
G F
GF
D001331:Automation; D015373:Bacterial Typing Techniques / Q000295:instrumentation* / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics; D005787:Gene Frequency; D005798:Genes, Bacterial; D005838:Genotype; D006801:Humans; D008589:Meningococcal Infections / Q000382:microbiology*; D009345:Neisseria meningitidis / Q000145:classification* / Q000235:genetics*; D010641:Phenotype; D018272:Porins / Q000235:genetics; D017422:Sequence Analysis, DNA
D004269:DNA, Bacterial; D018272:Porins; C057699:porin protein, Neisseria