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Rapid Identification and Susceptibility Testing of Uropathogenic Microbes via Immunosorbent ATP-Bioluminescence Assay on a Microfluidic Simulator for Antibiotic Therapy

Dong Tao, Zhao Xinyan
Analytical chemistry 2015 v.87 no.4 pp. 2410-2418
adenosine triphosphate, antibiotic resistance, antibiotics, antibodies, antibody specificity, antiseptics, bioluminescence, fiberglass, geometry, microorganisms, models, pathogen identification, pathogens, polypropylenes, therapeutics, urinary tract diseases, urine
The incorporation of pathogen identification with antimicrobial susceptibility testing (AST) was implemented on a concept microfluidic simulator, which is well suited for personalizing antibiotic treatment of urinary tract infections (UTIs). The microfluidic device employs a fiberglass membrane sandwiched between two polypropylene components, with capture antibodies immobilized on the membrane. The chambers in the microfluidic device share the same geometric distribution as the wells in a standard 384-well microplate, resulting in compatibility with common microplate readers. Thirteen types of common uropathogenic microbes were selected as the analytes in this study. The microbes can be specifically captured by various capture antibodies and then quantified via an ATP bioluminescence assay (ATP-BLA) either directly or after a variety of follow-up tests, including urine culture, antibiotic treatment, and personalized antibiotic therapy simulation. Owing to the design of the microfluidic device, as well as the antibody specificity and the ATP-BLA sensitivity, the simulator was proven to be able to identify UTI pathogen species in artificial urine samples within 20 min and to reliably and simultaneously verify the antiseptic effects of eight antibiotic drugs within 3–6 h. The measurement range of the device spreads from 1 × 10³ to 1 × 10⁵ cells/mL in urine samples. We envision that the medical simulator might be broadly employed in UTI treatment and could serve as a model for the diagnosis and treatment of other diseases.