Abstract Background Cefiderocol is among the latest generation of commercialized antibiotics against a large variety of MDR Gram-negative bacteria including carbapenem-resistant Enterobacterales and non-fermenters such as Pseudomonas aeruginosa and Acinetobacter baumannii. Cefiderocol susceptibility testing, a key element for implementing rapidly a cefiderocol-based treatment, might be still challenging. Objectives To develop a rapid culture-based test, Rapid Cefiderocol NP test, for the identification of cefiderocol resistance among MDR Enterobacterales. Methods The Rapid Cefiderocol NP test is based on glucose metabolization when bacterial growth occurs and the detection of bacterial growth in the presence of cefiderocol at 64 mg/L using iron-depleted CAMHB. Bacterial growth is visually detectable by a red-to-yellow colour change of red phenol, a pH indicator. A total of 74 clinical enterobacterial isolates from various clinical sources and of worldwide origin, among which 42 isolates were cefiderocol resistant, were used to evaluate the test performance. Results The sensitivity and specificity of the test were found to be 98% and 91%, respectively, by comparison with the reference broth microdilution (BMD) method. All positive results were obtained within 3 h after incubation at 35°C ± 2°C, that is a gain of time of ca. 18 h (1 day) compared with currently used techniques for susceptibility testing (BMD method). Conclusions This novel test is rapid, highly sensitive, specific, easily interpretable, and easy to implement in routine microbiology laboratories. Such a test may rapidly and accurately provide the information needed for the implementation of adequate cefiderocol-based treatment.