Automated web-based typing of Clostridioides difficile ribotypes via MALDI-TOF MS

Abstract Background Clostridioides difficile is a major cause of hospital-acquired diarrhea and a driver of nosocomial outbreaks, yet rapid, accurate ribotype identification remains challenging. We sought to develop a MALDI-TOF MS–based workflow coupled with machine learning to distinguish epidemic...

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Main Authors: Mario Blázquez-Sánchez, Alejandro Guerrero-López, Ana Candela, Albert Belenguer-Llorens, José Miguel Moreno, Carlos Sevilla-Salcedo, María Sánchez-Cueto, Manuel J. Arroyo, Mark Gutiérrez-Pareja, Vanessa Gómez-Verdejo, Pablo M. Olmos, Luis Mancera, Patricia Muñoz, Mercedes Marín, Luis Alcalá, David Rodríguez-Temporal, Belén Rodríguez-Sánchez
Format: Article
Language:English
Published: BMC 2025-07-01
Series:BMC Bioinformatics
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Online Access:https://doi.org/10.1186/s12859-025-06200-6
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Summary:Abstract Background Clostridioides difficile is a major cause of hospital-acquired diarrhea and a driver of nosocomial outbreaks, yet rapid, accurate ribotype identification remains challenging. We sought to develop a MALDI-TOF MS–based workflow coupled with machine learning to distinguish epidemic toxigenic ribotypes (RT027 and RT181) from other strains in real time. Results We analyzed MALDI-TOF spectra from 379 clinical isolates collected across ten Spanish hospitals and identified seven discriminant biomarker peaks. Two peaks (2463 and 4993 m/z) were uniquely associated with RT027, while combinations of five additional peaks reliably identified RT181. Our classifiers–implemented both in the commercial Clover MSDAS platform and the open-access AutoCdiff web tool–achieved up to 100% balanced accuracy in ribotype assignment and proved robust in real-time outbreak simulations. Conclusions This study demonstrates that MALDI-TOF MS combined with tailored machine learning can deliver rapid, high-precision ribotype identification for C. difficile. The freely available AutoCdiff models ( https://bacteria.id ) offer an immediately deployable solution for clinical laboratories, with the potential to enhance outbreak surveillance and control.
ISSN:1471-2105