Rapid Detection of Carbapenem-Producing and Enzyme Category in <i>Enterobacterales</i> through Ultraviolet Spectroscopy-Based Meropenem Hydrolysis Assessment

Carbapenem-producing Enterobacterales (CPE) pose a serious global health threat. This study aimed to develop a rapid and cost-effective ultraviolet (UV) spectrophotometry-based assay for detecting carbapenem resistance and identifying the carbapenemase enzyme category in Enterobacterales, thereby substantially reducing diagnostic turnaround times. The assay quantified Meropenem hydrolysis using ΔA = A300–A340 and was evaluated using 137 Enterobacterales strains. The hydrolysis level, measured by residual Meropenem after 3 h at 37 °C, was used to assess resistance. The A300–A340 method showed strong agreement with standard disk diffusion and VITEK-2 Compact results. Furthermore, the inhibitory effect of EDTA on Meropenem hydrolysis was tested on 79 CPE strains (35 MBL-producing and 44 non-MBL-producing) to differentiate carbapenemase categories, with PCR serving as the reference. A strong linear correlation (R2 = 0.9934) was observed between Meropenem concentration and ΔA. The assay demonstrated 98.54% accuracy in detecting Meropenem resistance using an 85% Meropenem retention threshold, and both the hydrolysis and inhibitor enhancement assays showed 97.47% specificity for distinguishing MBL from non-MBL producers. In conclusion, this UV spectrophotometry-based assay provides a rapid, accurate, and economical approach for detecting carbapenem resistance and classifying carbapenemase type, offering great potential for improving CPE infection management and guiding antibiotic therapy.