Direct Klebsiella pneumoniae Carbapenem Resistance and Carbapenemases\nGenotype Prediction by\nAl-MOF/TiO<sub>2</sub>@Au Cubic Heterostructures-Assisted Intact Bacterial\nCells Metabolic Analysis

Carbapenem-resistant Klebsiella pneumoniae (CRKP) infections pose a significant threat to human health. Fast\nand accurate prediction of K. pneumoniae carbapenem resistance and carbapenemase genotype is critical for\nguiding antibiotic treatment and reducing mortality rates. In this\nstudy, we present a novel method using Al-MOF/TiO<sub>2</sub>@Au cubic\nheterostructures for the metabolic analysis of intact bacterial cells,\nenabling rapid diagnosis of CRKP and its carbapenemases genotype.\nThe Al-MOF/TiO<sub>2</sub>@Au cubic composites display strong light\nabsorption and high surface area, facilitating the in situ effective\nextraction of metabolic fingerprints from intact bacterial cells.\nUtilizing this method, we rapidly and sensitively extracted metabolic\nfingerprints from 169 clinical isolates of K. pneumoniae obtained from patients. Machine learning analysis of the metabolic\nfingerprint changes successfully distinguishes CRKP from the sensitive\nstrains, achieving the high area under the curve (AUC) values of 1.00\nin both training and testing sets based on the 254 <i>m</i>/<i>z</i> features, respectively. Additionally, this platform\nenables rapid carbapenemase genotype discrimination of CRKP for precision\nantibiotic therapy. Our strategy holds great potential for swift diagnosis\nof CRKP and carbapenemase genotype discrimination, guiding effective\nmanagement of CRKP bacterial infections in both hospital and community\nsettings.