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Rapid one-step extraction method for the identification of molds using MALDI-TOF MS
Paul M Luethy1, Adrian M Zelazny1
1Microbiology Service, Department of Laboratory Medicine, Clinical Center, National Institutes of Health, Bethesda, MD, USA.
Diagnostic Microbiology and Infectious Disease
|February 19, 2018
Summary
A new rapid protein extraction method enhances fungal identification using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). This faster, reproducible technique improves accuracy and accessibility for clinical laboratories identifying molds.
Area of Science:
- Microbiology
- Analytical Chemistry
- Biotechnology
Background:
- Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a powerful tool for fungal identification.
- Widespread adoption of MALDI-TOF MS for mold identification is hindered by the lack of a simple, reproducible protein extraction protocol.
Purpose of the Study:
- To develop and validate a rapid, one-step protein extraction protocol for filamentous fungi.
- To improve the efficiency and reproducibility of MALDI-TOF MS-based mold identification.
Main Methods:
- A novel one-step extraction protocol using zirconia-silica beads and optimized extraction solutions was developed.
- Filamentous fungi were homogenized using a bead-based homogenizer, and supernatants were analyzed by MALDI-TOF MS.
- The rapid method was prospectively evaluated against a current routine protocol using 106 clinical mold isolates.
Main Results:
- The rapid method reduced hands-on extraction time to 5 minutes, a decrease of 30 minutes.
- Improved identification performance was observed, with acceptable scores (≥ 2.00) for 63.0% of isolates compared to 52.8% with the routine protocol.
- The rapid method demonstrated higher reproducibility between duplicate spots.
Conclusions:
- The developed rapid one-step protein extraction protocol is efficient and reproducible for MALDI-TOF MS analysis of clinical mold isolates.
- This method simplifies the adoption of MALDI-TOF MS for fungal identification in clinical laboratories.
- The protocol supports both batch and on-demand analysis, enhancing laboratory workflow flexibility.
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