Semiautomation of multilocus sequence typing for the characterization of clinical isolates of Neisseria meningitidis

The Scottish Meningococcus and Pneumococcus Reference Laboratory (SMPRL) pr ovides a national service for the laboratory confirmation of meningococcal and pneumococcal disease in Scotland. Part of this service includes the ser ogrouping of meningococcal isolates followed by typing and subtyping. The p rocedures for this are labor-intensive but important for the identification of linked cases and the surveillance of disease so that effective public h ealth measures can be taken. However, different strains of meningococci, su ch as those within the electrophoretic type 37 complex, occurring during ca se clusters of disease are now indistinguishable by current methods. The SM PRL has started using multilocus sequence typing (MLST) as a routine method for the characterization of isolates of Neisseria meningitidis. MLST produ ces nucleotide sequence data of seven housekeeping. genes providing results that are useful for public health management. However, the method is labor ious and time-consuming and therefore lends itself towards automation. The SMPRL therefore developed a semiautomated method for MLST using a 96-well f ormat liquid handler and an automated DNA sequencer. Semiautomated MLST is now provided as a reference service for Scotland. This work describes the m ethodology required for the characterization of N. meningitidis and highlig hts its usefulness for public health intervention.