In this section
@article{Gonzalo:2021:10.1016/j.cmi.2020.08.027,
author = {Gonzalo, X and Broda, A and Drobniewski, F and Larrouy-Maumus, G},
doi = {10.1016/j.cmi.2020.08.027},
journal = {Clinical Microbiology and Infection},
pages = {912.e1--912.e5},
title = {Performance of lipid fingerprint-based MALDI-ToF for the diagnosis of mycobacterial infections},
url = {http://dx.doi.org/10.1016/j.cmi.2020.08.027},
volume = {27},
year = {2021}
}
TY - JOUR
AB - ObjectivesBacterial diagnosis of mycobacteria is often challenging because of the variability of the sensitivity and specificity of the assay used, and it can be expensive to perform accurately. Although matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) has become the workhorse of clinical laboratories, the current MALDI methodology (which is based on cytosolic protein profiling) for mycobacteria is still challenging due to the number of steps involved (up to seven) and potential biosafety concerns. Knowing that mycobacteria produce surface-exposed species-specific lipids, we here hypothesized that the detection of those molecules could offer a rapid, reproducible and robust method for mycobacterial identification.MethodsWe evaluated the performance of an alternative methodology based on characterized species-specific lipid profiling of intact bacteria, without any sample preparation, by MALDI MS; it uses MALDI-time-of-flight (ToF) MS combined with a specific matrix (super-2,5-dihydroxybenzoic acid solubilized in an apolar solvent system) to analyse lipids of intact heat-inactivated mycobacteria. Cultured mycobacteria are heat-inactivated and loaded directly onto the MALDI target followed by addition of the matrix. Acquisition of the data is done in both positive and negative ion modes. Blinded studies were performed using 273 mycobacterial strains comprising both the Mycobacterium tuberculosis (Mtb) complex and non-tuberculous mycobacteria (NTMs) subcultured in Middlebrook 7H9 media supplemented with 10% OADC (oleic acid/dextrose/catalase) growth supplement and incubated for up to 2 weeks at 37°C.ResultsThe method we have developed is fast (<10 mins) and highly sensitive (<1000 bacteria required); 96.7% of the Mtb complex strains (204/211) were correctly assigned as MTB complex and 91.7% (22/24) NTM species were correctly assigned based only on intact bacteria species-specific lipid profiling by MALDI-ToF MS.ConclusionsIntact bacter
AU - Gonzalo,X
AU - Broda,A
AU - Drobniewski,F
AU - Larrouy-Maumus,G
DO - 10.1016/j.cmi.2020.08.027
EP - 1
PY - 2021///
SN - 1198-743X
SP - 912
TI - Performance of lipid fingerprint-based MALDI-ToF for the diagnosis of mycobacterial infections
T2 - Clinical Microbiology and Infection
UR - http://dx.doi.org/10.1016/j.cmi.2020.08.027
UR - https://www.sciencedirect.com/science/article/pii/S1198743X20305103?via%3Dihub
UR - http://hdl.handle.net/10044/1/82651
VL - 27
ER -
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