Tandem Bioluminescent Tests for Rapid Detection of Urinary Tract Infections and Antimicrobial Resistance Using a Portable Luminometer.

Urinary tract infections (UTIs) are among the most common bacterial infections in the world. Additionally, increasing antimicrobial resistance (AMR) further complicates the task of effective therapy for UTIs and, thus, presents a serious public health hazard. The standard diagnosis for UTI and AMR requires access to laboratory infrastructures, well-trained personnel, and long periods of processing time, all of which are factors increasing healthcare costs and inappropriate antibiotic treatment. Here, we designed bioluminescence-based tests for UTI and AMR detection that can be performed in tandem using a portable luminometer in ∼30 min. To date, no methods have been reported that can simultaneously detect both UTI and AMR using the same detection principle and portable device as described here, offering a significant advantage in streamlining testing. Our UTI test is based on whole-cell bioluminescence assay, and AMR detection employs a modified ATP-based bioluminescence detection. Our test can detect UTI in 5 min at a bacterial level as low as 20,000 CFU/mL, which meets the clinically significant threshold of 100,000 CFU/mL. Antibiotic resistance was monitored using a modified ATP-based bioluminescence test within 30 min. The test showed no interference from potential urinary components, such as glucose, urea, commensal flora, or pH variations. A clinical sample (n = 8) analysis using these tests showed 100% accuracy in diagnosing UTI and AMR. These tandem tests using a portable bioluminescent reader could potentially minimize delays in diagnosis to allow immediate clinical decisions and optimization of treatment with antibiotics in clinical or community health settings.