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. 2022 Jul 12;4(7):acmi000372.
doi: 10.1099/acmi.0.000372. eCollection 2022 Aug.

Rapid identification of clinically interesting methanogens using an improved MALDI-TOF-MS assay

Cheick Oumar Guindo  1   2 , Lynda Amir  1   2 , Carine Couderc  3 , Michel Drancourt  1   2   3 , Ghiles Grine  1   2
Affiliations

Rapid identification of clinically interesting methanogens using an improved MALDI-TOF-MS assay

Cheick Oumar Guindo et al. Access Microbiol. .

Abstract

Methanogens, the archaea uniquely detoxifying fermentative hydrogen into methane in the digestive tract, are increasingly being detected in pathology situations, rendering their rapid identification mandatory. We improved the experimental protocol to identify broth-cultured methanogens by matrix-assisted laser desorption time-of-flight MS (MALDI-TOF-MS). A database incorporating 34 reference spectra derived from 16 methanogen reference strains representative of eight species supported further identification of 21 Methanobrevibacter smithii and 14 Methanobrevibacter oralis isolates broth-cultured from human stool and oral fluid, respectively, with scores >2. In addition, MALDI-TOF-MS differentiated five Methanobrevibacter smithii genotypes incorporated in the study. The data reported here found MALDI-TOF-MS as a first-line identification method for methanogens recovered from microbiota and clinical samples.

Keywords: MALDI-TOF-MS; clinical microbiology; culture; identification; methanogens; multispacer sequence typing.

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Conflict of interest statement

The authors declare that there are no conflicts of interest.

Figures

Fig. 1.
Fig. 1.
Comparison of the MALDI-TOF-MS spectra of the 16 reference methanogen strains using MALDI Biotyper version 4.1.80 software (Bruker Daltonics). Normalized gel view representation of the MS profile intensity of the 16 reference methanogenic strains used in this study. a.u., Arbitrary units; m/z, mass-to-charge ratio.
Fig. 2.
Fig. 2.
Representative MS profiles of the 16 methanogen reference strains studied using FlexAnalysis software. (1) MS profiles of Methanobacterium beijingense CSURP9638. (2) MS profiles of Methanobrevibacter arboriphilicus CSURP9635. (3) MS profiles of M. luminyensis CSURP9636. (4) MS profiles of Methanosarcina barkeri CSURQ5732. (5) MS profiles of Methanobrevibacter smithii CSURP5922. (6) MS profiles of Methanobrevibacter smithii CSURP5816. (7) MS profiles of Methanobrevibacter smithii CSURQ5493. (8) MS profiles of Methanobrevibacter smithii CSURQ5497. (9) MS profiles of Methanobrevibacter smithii CSURQ5501. (10) MS profiles of Methanosarcina mazei CSURP9637. (11) MS profiles of Methanosphaera stadtmanae CSURP9634. (12) MS profiles of Methanobrevibacter oralis CSURP5701. (13) MS profiles of Methanobrevibacter oralis CSURQ5479. (14) MS profiles of Methanobrevibacter oralis CSURQ5481. (15) MS profiles of Methanobrevibacter oralis CSURQ5483. (16) MS profiles of Methanobrevibacter oralis CSURQ5485. The 16 methanogen reference strains are presented in duplicate to demonstrate the reproducibility of MS profiles. a.u., Arbitrary units; m/z, mass-to-charge ratio.
Fig. 3.
Fig. 3.
Negative controls used in the MALDI-TOF-MS experiment. Non-inoculated media are in blue and matrix solution in red. a.u., Arbitrary units; m/z, mass-to-charge ratio.
See this image and copyright information in PMC

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