In this study, we evaluate the performance of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for rapid detection of carbapenemase activity in Enterobacterales in clinical microbiology laboratories during a multicenter networking validation study. The study was divided into three different stages: "software design," "intercenter evaluation," and "clinical validation." First, a standardized procedure with an online software for data analysis was designed. Carbapenem resistance was detected by measuring imipenem hydrolysis and the results were automatically interpreted using the Clover MS data analysis software (Clover BioSoft, Spain). Second, a series of 74 genotypically characterized Enterobacterales (46 carbapenemase-producers and 28 non carbapenemase-producers) were analyzed in 8 international centers to ensure the reproducibility of the method. Finally, the methodology was evaluated independently in all centers during a 2-month period and results were compared with the reference standard for carbapenemase detection used in each center. The overall agreement rate relative to the reference method for carbapenemase resistance detection in clinical samples was 92.5%. The sensitivity was 93.9% and the specificity, 100%. Results were obtained within 60 min and accuracy ranged from 83.3 to 100% among the different centers. Further, our results demonstrate that MALDI-TOF MS is an outstanding tool for rapid detection of carbapenemase activity in Enterobacterales in clinical microbiology laboratories. The use of a simple in-house procedure with online software allows routine screening of carbapenemases in diagnostics, thereby facilitating early and appropriate antimicrobial therapy.

Clover BioSoft Granada Spain

Departamento de Química Agrícola Edafología y Microbiología Universidad de Córdoba Córdoba Spain

Department of Microbiology Faculty of Medicine University Hospital in Pilsen Charles University Pilsen Czechia

Instituto Maimónides de Investigación Biomédica de Córdoba Córdoba Spain

Service de Microbiologie Cliniques Universitaires Saint Luc Brussels Belgium

Servicio de Análisis Clínicos Complejo Hospitalario Universitario A Coruña A Coruña Spain

Servicio de Microbiología Hospital General Universitario Gregorio Marañón Madrid Spain

Servicio de Microbiología Hospital Universitario Central de Asturias Instituto de Investigación Sanitaria del Principado de Asturias Oviedo Spain

Servicio de Microbiología Hospital Universitario Son Espases Red Española de Investigación en Patología Infecciosa CIBER de Enfermedades Infecciosas Instituto de Salud Carlos 3 Palma Spain

Servicio de Microbiología Red Española de Investigación en Patología Infecciosa Hospital Universitario Ramón y Cajal Instituto Ramón y Cajal de Investigación Sanitaria CIBER de Enfermedades Infecciosas Instituto de Salud Carlos 3 Madrid Spain

Servicio de Microbiología Red Española de Investigación en Patología Infecciosa Instituto de Investigación Biomédica da Coruña CIBER de Enfermedades Infecciosas Instituto de Salud Carlos 3 Complejo Hospitalario Universitario A Coruña A Coruña Spain

Unidad Clínica de Enfermedades Infecciosas y Microbiología Clínica CSIC Red Española de Investigación en Patología Infecciosa Hospital Universitario Virgen Macarena Instituto de Biomedicina de Sevilla CIBER de Enfermedades Infecciosas Instituto de Salud Carlos 3 Universidad de Sevilla Seville Spain

Unidad de Gestión Clínica de Microbiología Red Española de Investigación en Patología Infecciosa Hospital Universitario Reina Sofía CIBER de Enfermedades Infecciosas Instituto de Salud Carlos 3 Córdoba Spain

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