Nadine Perrot (Presenter)
R&D Microbiology Department, bioMérieux S.A.
Authorship: N. Perrot (1), P. Broyer (2), J. Blaze (2), C. Bailey (3), J.M. Baumlin (4), J.P. Charrier (5)
(1) R&D Microbiology Department, bioMérieux S.A., La Balme Les Grottes FR, (2) Innovation Unit, bioMérieux S.A., Grenoble FR, (3) R&D System Development, bioMérieux Inc. , St Louis USA, (4) R&D Product Development, Arjowiggins Creative Papers, Boulogne-Billancourt FR, (5) Microbiology Research Department, bioMérieux S.A., Marcy l'Etoile FR
MALDI-TOF MS is commonly used as a tool for the rapid identification of microorganisms in clinical and industrial fields. Samples are classically loaded using either disposable plastic targets or reusable metallic slides. To reduce the environmental footprint of MALDI-TOF applications, the use of a low cost, “green target” made with paper substratum, was proposed. A proof of concept was realized with excellent results using different types of paper with a metallized side as new sample supports for microorganism identification and molecule detection (peptides and protein up to 46kDa).
MALDI-TOF MS is commonly used as a tool for the rapid identification of microorganisms in clinical and industrial fields. Samples are classically loaded using either disposable plastic targets or reusable metallic slides. Before new sample deposition, the latter need time-consuming cleaning steps with solvents. To reduce the environmental footprint of MALDI-TOF applications, the use of a low cost, “green target” made with paper substratum, was proposed.
Different types of paper with a metallized side were tested as new sample supports for microorganism identification and molecule detection by MALDI-TOF. The identification was performed on 22 strains (8 Gram positives, 11 Gram negatives, 3 yeasts). Pure colonies were manually streaked on the paper slide, 1 µl of CHCA matrix was added (preceded by 0.5 µl of formic acid for yeast). Acquisition and identification was performed using VITEK-MS RUO mode (launchpad1.4.2/ SARAMIS). Peptides, proteins, and oligosaccharides were detected using different matrixes (DHB, CHCA, SA, DHB-py, DHB-mi) and appropriate mass windows.
Microorganism identification on paper targets were as good as identification on commercialized VITEK-MS disposable targets and were 100% correct for 8 Gram positives, 11 Gram negatives and 3 yeasts. Identification probabilities were similar with averages of 98,4%, 96,5%, and 98% for the reference, the smooth, and the structured paper, respectively. MS spectra were also comparable: similar resolution, number of peaks, and dynamic range. All the peptides, proteins (up to 46 kDa), and oligosaccharides tested with the different matrices were detected on the smooth paper with the same quality as on the reference target.
Conclusions & Discussion
Disposable MALDI-TOF targets, environmentally friendly, are possible using paper (cellulose substratum) with a metallic side. Excellent performances were obtained for microorganism identification and detection of oligosaccharides, peptides, and proteins, using wide mass range windows. Paper targets have potential for mass production at low cost per target.
References & Acknowledgements:
IP Royalty: no
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