Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the identification of Burkholderia pseudomallei from Asia and Australia and differentiation between Burkholderia species

Vichaya Suttisunhakul, Apinya Pumpuang, Peeraya Ekchariyawat, Vanaporn Wuthiekanun, Mindy Glass Elrod, Paul Turner, Paul Turner, Bart J. Currie, Bart J. Currie, Rattanaphone Phetsouvanh, Rattanaphone Phetsouvanh, David A B Dance, David A B Dance, David A B Dance, Direk Limmathurotsakul, Direk Limmathurotsakul, Sharon J Peacock, Narisara Chantratita, Narisara Chantratita

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Abstract

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is increasingly used for rapid bacterial identification. Studies of Burkholderia pseudomallei identification have involved small isolate numbers drawn from a restricted geographic region. There is a need to expand the reference database and evaluate B. pseudomallei from a wider geographic distribution that more fully captures the extensive genetic diversity of this species. Here, we describe the evaluation of over 650 isolates. Main spectral profiles (MSP) for 26 isolates of B. pseudomallei (N = 5) and other Burkholderia species (N = 21) were added to the Biotyper database. MALDI-TOF MS was then performed on 581 B. pseudomallei, 19 B. mallei, 6 B. thailandensis and 23 isolates representing a range of other bacterial species. B. pseudomallei originated from northeast and east Thailand (N = 524), Laos (N = 12), Cambodia (N = 14), Hong Kong (N = 4) and Australia (N = 27). All 581 B. pseudomallei were correctly identified, with 100% sensitivity and specificity. Accurate identification required a minimum inoculum of 5 x 107 CFU/ml, and identification could be performed on spiked blood cultures after 24 hours of incubation. Comparison between a dendrogram constructed from MALDI-TOF MS main spectrum profiles and a phylogenetic tree based on recA gene sequencing demonstrated that MALDI-TOF MS distinguished between B. pseudomallei and B. mallei, while the recA tree did not. MALDI-TOF MS is an accurate method for the identification of B. pseudomallei, and discriminates between this and other related Burkholderia species.
Original languageEnglish
Article numbere0175294
Pages (from-to)1-16
Number of pages16
JournalPLoS One
Volume12
Issue number4
DOIs
Publication statusPublished - 1 Apr 2017

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    Suttisunhakul, V., Pumpuang, A., Ekchariyawat, P., Wuthiekanun, V., Elrod, M. G., Turner, P., Turner, P., Currie, B. J., Currie, B. J., Phetsouvanh, R., Phetsouvanh, R., Dance, D. A. B., Dance, D. A. B., Dance, D. A. B., Limmathurotsakul, D., Limmathurotsakul, D., Peacock, S. J., Chantratita, N., & Chantratita, N. (2017). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the identification of Burkholderia pseudomallei from Asia and Australia and differentiation between Burkholderia species. PLoS One, 12(4), 1-16. [e0175294]. https://doi.org/10.1371/journal.pone.0175294