Multilocus Sequence Typing of Streptococcus pneumoniae by Use of Mass Spectrometry
Eileen M Dunne, Eng Kok Ong, Ralf J Moser, Peter M Siba, Suparat Phuanukoonnon, Andrew R Greenhill, Roy M Robins-Browne, E Kim Mulholland, Catherine Satzke
Journal of Clinical Microbiology | AMER SOC MICROBIOLOGY | Published : 2011
Multilocus sequence typing (MLST) is an important tool for the global surveillance of bacterial pathogens that is performed by comparing the sequences of designated housekeeping genes. We developed and tested a novel mass spectrometry-based method for MLST of Streptococcus pneumoniae. PCR amplicons were subjected to in vitro transcription and base-specific cleavage, followed by analysis of the resultant fragments by using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Comparison of the cleavage fragment peak patterns to a reference sequence set permitted automated identification of alleles. Validation experiments using 29 isolates of S. pneumonia..View full abstract
This work was supported by funding from the Murdoch Childrens Research Institute, the American Australian Association, and the Victorian Government's Operational Infrastructure Support Program. We acknowledge the use of the pneumococcal MLST database, which is located at Imperial College London and is funded by the Wellcome Trust.