Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry-Based Functional Assay for Rapid Detection of Resistance against β-Lactam Antibiotics

Resistance against beta-lactam antibiotics is a growing challenge for managing severe bacterial infections. The rapid and cost-efficient determination of beta-lactam resistance is an important prerequisite for the choice of an adequate antibiotic therapy. beta-Lactam resistance is based mainly on the expression/overexpression of beta-lactamases, which destroy the central beta-lactam ring of these drugs by hydrolysis. Hydrolysis corresponds to a mass shift of + 18 Da, which can be easily detected by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Therefore, a MALDI-TOF MS-based assay was set up to investigate different enterobacteria for resistance against different beta-lactam antibiotics: ampicillin, piperacillin, cefotaxime, ceftazidime, ertapenem, imipenem, and meropenem. beta-Lactamases are enzymes that have a high turnover rate. Therefore, hydrolysis can be detected by MALDI-TOF MS already after a few hours of incubation of the bacteria to be tested with the given antibiotic. The comparison of the MS-derived data with the data from the routine procedure revealed identical classification of the bacteria according to sensitivity and resistance. The MALDI-TOF MS-based assay delivers the results on the same day. The approved routine procedures require at least an additional overnight incubation.