Implementation of MALDI-TOF Mass Spectrometry and Peak Analysis: Application to the Discrimination of &lt;em&gt;Cryptococcus&lt;/em&gt; Species and Their Interspecies Hybrids

Zvezdánova, Margarita Estreya; Arroyo, Manuel J.; Méndez, Gema; Guinea, Jesús; Mancera, Luis; Muñóz, Patricia; Rodríguez‐Sánchez, Belén; Escribano, Pilar

doi:10.20944/preprints202007.0423.v1

Cited by 3 publications

References 26 publications

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Rapid and reproducible MALDI-TOF-based method for detection Vancomycin-resistant Enterococcus faecium using classifying algorithms

Candela

Arroyo

Sánchez-Molleda

et al. 2021

Preprint

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Vancomycin-resistant Enterococcus faecium has become a health threat over the last 20 years due to its ability to rapidly spread and cause outbreaks in hospital settings. Although MALDI-TOF MS has already demonstrated its usefulness for accurate identification of E. faecium, its implementation for antimicrobial resistance detection is still under evaluation. The reproducibility of MALDI-TOF MS for peak analysis and its performance for correct discrimination of vancomycin susceptible isolates (VSE) from those hosting the VanA and VanB resistance mechanisms was evaluated in this study. For the first goal, intra-spot, inter-spot -technical- and inter-day -biological- reproducibility was assayed. The capability of MALDI-TOF to discriminate VSE isolates from VanA VRE and VanB VRE strains was carried out on protein spectra from 178 E. faecium unique clinical isolates −92 VSE, 31 VanA VRE, 55 VanB VRE-, processed with Clover MS Data Analysis software. Unsupervised (Principal Component Analysis –PCA-) and supervised algorithms (Support Vector Machine -SVM-, Random Forest -RF- and Partial Least Squares-Discriminant Analysis -PLS-DA-) were applied. The reproducibility assay showed lower variability for normalized data (p<0.0001) and for the peaks within the 3000-9000 m/z range. Besides, 80.9%, 79.21% and 77.53% VSE vs VRE (VanA + VanB) discrimination was achieved by applying SVM, RF and PLS-DA, respectively. Correct differentiation of VanA from VanB VRE isolates was obtained by SVM in 86.65% cases. The implementation MALDI-TOF MS and peak analysis could represent a rapid and effective tool for VRE screening. However, further improvements are needed to increase the accuracy of this approach.

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Rapid and reproducible MALDI-TOF-based method for detection Vancomycin-resistant Enterococcus faecium using classifying algorithms

Candela

Arroyo

Sánchez-Molleda

et al. 2021

Preprint

Self Cite

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Rapid and Reproducible MALDI-TOF-Based Method for the Detection of Vancomycin-Resistant Enterococcus faecium Using Classifying Algorithms

et al. 2022

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Vancomycin-resistant Enterococcus faecium represents a health threat due to its ability to spread and cause outbreaks. MALDI-TOF MS has demonstrated its usefulness for E. faecium identification, but its implementation for antimicrobial resistance detection is still under evaluation. This study assesses the repeatability of MALDI-TOF MS for peak analysis and its performance in the discrimination of vancomycin-susceptible (VSE) from vancomycin-resistant isolates (VRE). The study was carried out on protein spectra from 178 E. faecium unique clinical isolates—92 VSE, 31 VanA VRE, 55 VanB VRE-, processed with Clover MS Data Analysis software. Technical and biological repeatability were assayed. Unsupervised (principal component analysis, (PCA)) and supervised algorithms (support vector machine (SVM), random forest (RF) and partial least squares–discriminant analysis (PLS-DA)) were applied. The repeatability assay was performed with 18 peaks common to VSE and VRE with intensities above 1.0% of the maximum peak intensity. It showed lower variability for normalized data and for the peaks within the 3000–9000 m/z range. It was found that 80.9%, 79.2% and 77.5% VSE vs. VRE discrimination was achieved by applying SVM, RF and PLS-DA, respectively. Correct internal differentiation of VanA from VanB VRE isolates was obtained by SVM in 86.6% cases. The implementation of MALDI-TOF MS and peak analysis could represent a rapid and effective tool for VRE screening. However, further improvements are needed to increase the accuracy of this approach.

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MALDI-TOF MS in a Medical Mycology Laboratory: On Stage and Backstage

et al. 2021

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The implementation of MALDI-TOF MS in medical microbiology laboratories has revolutionized practices and significantly reduced turnaround times of identification processes. However, although bacteriology quickly benefited from the contributions of this technique, adjustments were necessary to accommodate the specific characteristics of fungi. MALDI-TOF MS is now an indispensable tool in clinical mycology laboratories, both for the identification of yeasts and filamentous fungi, and other innovative uses are gradually emerging. Based on the practical experience of our medical mycology laboratory, this review will present the current uses of MALDI-TOF MS and the adaptations we implemented, to allow their practical execution in a daily routine. We will also introduce some less mainstream applications, like those for fungemia, or even still under development, as is the case for the determination of sensitivity to antifungal agents or typing methods.

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Implementation of MALDI-TOF Mass Spectrometry and Peak Analysis: Application to the Discrimination of <em>Cryptococcus</em> Species and Their Interspecies Hybrids

Cited by 3 publications

References 26 publications

Rapid and reproducible MALDI-TOF-based method for detection Vancomycin-resistant Enterococcus faecium using classifying algorithms

Rapid and reproducible MALDI-TOF-based method for detection Vancomycin-resistant Enterococcus faecium using classifying algorithms

Rapid and Reproducible MALDI-TOF-Based Method for the Detection of Vancomycin-Resistant Enterococcus faecium Using Classifying Algorithms

MALDI-TOF MS in a Medical Mycology Laboratory: On Stage and Backstage

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