Rapid Identification and Classification of Pathogens That Produce Carbapenemases and Cephalosporinases with a...

Area of Science:

• Biomedical Engineering
• Clinical Diagnostics
• Microbiology

Background:

• Bacterial infections producing β-lactamases (BLs) pose significant hospital challenges.
• Current detection methods require up to 72 hours of culturing, delaying critical antibiotic treatment.
• Rapid identification of BL-producing pathogens is essential for effective clinical management.

Purpose of the Study:

• To develop a rapid, culture-independent multisensor for identifying bacterial β-lactamases.
• To detect specific enzyme types like carbapenemases, cephalosporinases, metallo-β-lactamases, extended-spectrum β-lactamases (ESBLs), and AmpC enzymes.
• To provide a diagnostic tool that guides initial antibiotic therapy.

Main Methods:

• A paper-based multisensor platform was designed utilizing a cellulose scaffold for bacterial preconcentration.
• Twelve parallel assays were performed to detect various β-lactamase activities.
• Colorimetric detection and image processing quantified enzyme activity based on antibiotic hydrolysis and inhibition patterns.

Main Results:

• The multisensor successfully identified pathogens with different β-lactamases above infectious dose thresholds within 90 minutes.
• The system demonstrated specificity in detecting various enzyme classes, including metallo-β-lactamases, ESBLs, and AmpC.
• Validation for diagnosing urinary tract infections confirmed the prototype's efficacy.

Conclusions:

• The developed paper-based multisensor offers a rapid, culture-free method for detecting β-lactamase-producing bacteria.
• Its inexpensive nature and simple colorimetric readout are suitable for developing automated diagnostic platforms.
• This technology promises streamlined clinical diagnosis and improved patient outcomes for bacterial infections.