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Multilocus sequence typing of Streptococcus pneumoniae by use of mass spectrometry

Dunne, Eileen M., Ong, Eng Kok, Moser, Ralp J., Siba, Peter M., Phuanukoonnon, Suparat, Greenhill, Andrew R., Robins-Browne, Roy M., Mulholland, E. Kim and Satzke, Catherine (2011). Multilocus sequence typing of Streptococcus pneumoniae by use of mass spectrometry. Journal of Clinical Microbiology,49(11):3756-3760.

Document type: Journal Article
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IRMA ID kmckayxPUB47
Title Multilocus sequence typing of Streptococcus pneumoniae by use of mass spectrometry
Author Dunne, Eileen M.
Ong, Eng Kok
Moser, Ralp J.
Siba, Peter M.
Phuanukoonnon, Suparat
Greenhill, Andrew R.
Robins-Browne, Roy M.
Mulholland, E. Kim
Satzke, Catherine
Journal Name Journal of Clinical Microbiology
Publication Date 2011
Volume Number 49
Issue Number 11
ISSN 0095-1137   (check CDU catalogue open catalogue search in new window)
Start Page 3756
End Page 3760
Total Pages 5
Place of Publication Washington, United States
Publisher American Society for Microbiology
HERDC Category C1 - Journal Article (DIISR)
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 MALDI-TOF 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.

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