Rapid Dynamic Surface-Enhanced Raman Spectroscopy Detection of Amoxicillin-Mediated Morphological Changes in a Pathogen for Diagnosis of Clinical Urine Samples

The swift and stable detection of pathogens in urine samples holds significant implications for the immediate clinical diagnosis and treatment of urinary tract infections (UTIs). In this study, we propose a detection strategy utilizing a hybrid substrate composed of graphene oxide (GO) and silver nanoparticles (Ag NPs) for the detection of pathogens subjected to amoxicillin-mediated (amo-mediated) treatment. This strategy employs dynamic surface-enhanced Raman spectroscopy (D-SERS) for stable and rapid detection, capturing signal variations induced by amo-mediated changes in pathogen morphology. During the 5 min D-SERS detection time window, stable SERS signals were detected for three types of pathogens and four types of pathogens were successfully distinguished using principal component analysis (PCA). In comparison to conventional nanosubstrates, the GO/Ag NP hybrid substrate exhibits outstanding stability and enhancement effects. This approach enables the dual detection of the pathogen cell structure and metabolites, facilitating specific identification of pathogens in the urinary tract, with a detection limit for Escherichia coli reaching 1 × 10⁴ colony-forming units (CFU)/mL, meeting the clinical microbiology laboratory diagnostic standards for UTIs (10⁵ CFU/mL). Testing of 188 clinically collected urine samples using this strategy yielded a sensitivity (SENS) of 86.4% and a specificity (SPC) of 89.7%. This introduces a novel method for diagnosing UTIs, offering broad applications in the field of clinical pathogen detection.