Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

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

Version 1 : Received: 17 July 2020 / Approved: 19 July 2020 / Online: 19 July 2020 (19:07:20 CEST)

How to cite: Zvezdanova, M.E.; Arroyo, M.J.; Méndez, G.; Guinea, J.; Mancera, L.; Muñoz, P.; Rodríguez-Sánchez, B.; Escribano, P. Implementation of MALDI-TOF Mass Spectrometry and Peak Analysis: Application to the Discrimination of Cryptococcus Species and Their Interspecies Hybrids. Preprints 2020, 2020070423 (doi: 10.20944/preprints202007.0423.v1). Zvezdanova, M.E.; Arroyo, M.J.; Méndez, G.; Guinea, J.; Mancera, L.; Muñoz, P.; Rodríguez-Sánchez, B.; Escribano, P. Implementation of MALDI-TOF Mass Spectrometry and Peak Analysis: Application to the Discrimination of Cryptococcus Species and Their Interspecies Hybrids. Preprints 2020, 2020070423 (doi: 10.20944/preprints202007.0423.v1).

Abstract

MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization Time-of Flight) is a type of mass spectrometry (MS) that has been widely implemented for the rapid identification of microorganisms over the last decade. The accuracy and flexibility of this method has encouraged researchers to implement the analysis of protein spectra obtained by MALDI-TOF for the discrimination of close-related species and bacterial typing. In this study, a standardized methodology based on the detection of species-specific protein peaks from the spectra obtained with MALDI-TOF is described. The methodology was applied to a collection of Cryptococcus spp. (n=70) previously characterized by Amplified Fragment Length Polymorphism (AFLP) and sequencing of the ITS1-5.8S-ITS2 region. An expanded ad-hoc database was also built for their discrimination with MALDI-TOF. This approach did not allow the discrimination of the interspecies hybrids. However, the performance of peak analysis with the application of the PLS-DA and SVM algorithms in a two-step analysis allowed 96.95% and 96.55% correct discrimination of C. neoformans from the interspecies hybrids, respectively. Besides, PCA analysis prior to SVM provided 98.45% correct discrimination of the 3 analyzed species in a one-step analysis. The method is cost-efficient, rapid and user-friendly. The procedure can also be automatized for an optimized implementation in the laboratory routine.

Subject Areas

Cryptococcus spp; MALDI-TOF MS; peak analysis; in-house library; hierarchical clustering

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