Comparison of a multiplexed MassARRAY system with real-time allele-specific PCR technology for genotyping of methicillin-resistant Staphylococcus aureus

Syrmis, M. W., Moser, R. J., Whiley, D. M., Vaska, V., Coombs, G. W., Nissen, M. D., Sloots, T. P. and Nimmo, G. R. (2011) Comparison of a multiplexed MassARRAY system with real-time allele-specific PCR technology for genotyping of methicillin-resistant Staphylococcus aureus. Clinical Microbiology and Infection, 17 12: 1804-1810. doi:10.1111/j.1469-0691.2011.03521.x


Author Syrmis, M. W.
Moser, R. J.
Whiley, D. M.
Vaska, V.
Coombs, G. W.
Nissen, M. D.
Sloots, T. P.
Nimmo, G. R.
Title Comparison of a multiplexed MassARRAY system with real-time allele-specific PCR technology for genotyping of methicillin-resistant Staphylococcus aureus
Formatted title
Comparison of a multiplexed MassARRAY system with real-time allele-specific PCR technology for genotyping of methicillin-resistant Staphylococcus aureus
Journal name Clinical Microbiology and Infection   Check publisher's open access policy
ISSN 1198-743X
Publication date 2011-12-01
Sub-type Article (original research)
DOI 10.1111/j.1469-0691.2011.03521.x
Volume 17
Issue 12
Start page 1804
End page 1810
Total pages 7
Place of publication Oxford, England, U.K.
Publisher Wiley-Blackwell Publishing
Collection year 2012
Language eng
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.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 22 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 28 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Wed, 19 Oct 2011, 19:28:30 EST by Melanie Syrmis on behalf of Clinical Medical Virology Centre