Electrogenerated Chemiluminescence Biosensor Based on Functionalized Two-Dimensional Metal-Organic Frameworks for Bacterial Detection and Antimicrobial Susceptibility Assays.

Abstract

The emergence of antibiotic resistance has prompted the development of rapid antimicrobial susceptibility testing (AST) technologies to guide antibiotic prescription. A novel electrochemiluminescence (ECL) biosensor developed can quantitatively measure the binding between the lectin and lipopolysaccharide (LPS) on Gram-negative bacteria for bacterial determination and to characterize the antimicrobial activities of β-lactam and non-β-lactam antibiotics to normal and antibiotic-resistant bacteria. The biosensor utilizes ruthenium complex tagged concanavalin A (Ru-Con A) coated on NH2-MIL-53(Al) interface for LPS binding measurements. The decreased ECL signal obtained was directly proportional to increasing Escherichia coli (E. coli) BL21 concentrations. The sensitivity displayed logarithmic dependence in the range of (50-5.0) × 104 cells/mL, with a detection limit of 16 cells/mL. The minimum inhibitory concentration (MIC) values of antibiotics for normal E. coli BL21 were 0.02-0.2, 2-4, 0.002-0.02, and 0.2-1 mg/L for levofloxacin hydrochloride (LVX), tetracycline (TCY), imipenem (IPM), and cefpirome (CPO), respectively. The increased MIC values (8-16 and 4 mg/L for IMP and CPO, respectively) in New Delhi metallo-β-lactamase-1 expressing E. coli BL21 (NDM-1-E. coli BL21) indicated greater resistance to β-lactams in NDM-1-E. coli BL21 compared with normal E. coli BL21. Therefore, the changed ECL signal because of binding between LPS with the lectin has a relation with the type of antibiotic and bacterial strains, making the ECL biosensor promote clinical practicability and facilitate antibiotic stewardship.