Journal article
Comparison of a multiplexed MassARRAY system with real-time allele-specific PCR technology for genotyping of methicillin-resistant Staphylococcus aureus
Clinical Microbiology and Infection, Vol.17(12), pp.1804-1810
2011
Abstract
The Sequenom MassARRAY iPLEX single-nucleotide polymorphism (SNP) typing platform uses matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) coupled with single-base extension PCR for high-throughput multiplex SNP detection. In this study, we investigated the use of iPLEX MassARRAY technology for methicillin-resistant Staphylococcus aureus (MRSA) genotyping. A 16-plex MassARRAY iPLEX GOLD assay (MRSA-iPLEX) was developed that targets a set of informative SNPs and binary genes for MRSA characterization. The method was evaluated with 147 MRSA isolates, and the results were compared with those of an established SYBR Green-based real-time PCR system utilizing the same SNP-binary markers. A total of 2352 markers belonging to 44 SNP-binary profiles were analysed by both real-time PCR and MRSA-iPLEX. With real-time PCR as the reference standard, MRSA-iPLEX correctly assigned 2298 of the 2352 (97.7%) markers. Sequence variation in the MRSA-iPLEX primer targets accounted for the majority of MRSA-iPLEX erroneous results, highlighting the importance of primer target selection. MRSA-iPLEX provided optimal throughput for MRSA genotyping, and was, on a reagent basis, more cost-effective than the real-time PCR methods. The 16-plex MRSA-iPLEX is a suitable alternative to SYBR Green-based real-time PCR typing of major sequence types and clonal complexes of MRSA.
Details
- Title
- Comparison of a multiplexed MassARRAY system with real-time allele-specific PCR technology for genotyping of methicillin-resistant Staphylococcus aureus
- Authors/Creators
- M.W. Syrmis (Author/Creator)R.J. Moser (Author/Creator)D.M. Whiley (Author/Creator)V. Vaska (Author/Creator)G.W. Coombs (Author/Creator)M.D. Nissen (Author/Creator)T.P. Sloots (Author/Creator)G.R. Nimmo (Author/Creator)
- Publication Details
- Clinical Microbiology and Infection, Vol.17(12), pp.1804-1810
- Publisher
- Elsevier
- Identifiers
- 991005540521307891
- Copyright
- © 2011 The Authors. Clinical Microbiology and Infection
- Murdoch Affiliation
- Murdoch University
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- Domestic collaboration
- Citation topics
- 1 Clinical & Life Sciences
- 1.23 Antibiotics & Antimicrobials
- 1.23.2108 Microbial Diagnostics
- Web Of Science research areas
- Infectious Diseases
- Microbiology
- ESI research areas
- Immunology