Multilocus Sequence Typing of Streptococcus pneumoniae by Use of mass Spectrometry

Eileen Dunne, Eng Kok Ong, Ralp Moser, Peter Siba, Suparat Phuanukoonnon, Andrew Greenhill, Roy M Robins-Browne, Edward (Kim) MULHOLLAND, Catherine Satzke

    Research output: Contribution to journalArticle

    Abstract

    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. pneumoniae revealed that the results of MALDI-TOF MS MLST matched those obtained by traditional sequence-based MLST for 99% of alleles and that the MALDITOF MS method accurately identified two single-nucleotide variations. The MADLI-TOF MS method was then used for MLST analysis of 43 S. pneumoniae isolates from Papua New Guinean children. The majority of the isolates present in this population were not clonal and contained seven new alleles and 30 previously unreported sequence types. Copyright � 2011, American Society for Microbiology. All Rights Reserved.
    Original languageEnglish
    Pages (from-to)3756-3760
    Number of pages5
    JournalJournal of Clinical Microbiology
    Volume49
    Issue number11
    DOIs
    Publication statusPublished - 2011

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    Dunne, E., Ong, E. K., Moser, R., Siba, P., Phuanukoonnon, S., Greenhill, A., Robins-Browne, R. M., MULHOLLAND, E. K., & Satzke, C. (2011). Multilocus Sequence Typing of Streptococcus pneumoniae by Use of mass Spectrometry. Journal of Clinical Microbiology, 49(11), 3756-3760. https://doi.org/10.1128/JCM.05113-11