Microfluidic droplet-in-oil partitioning device for rapid phenotypic ast for Neisseria gonorrhoeae

Accurate and timely antimicrobial susceptibility testing (AST) is critical for managing gonorrhea infections. Current gold standard AST requires several days of turnaround time, which leads to compromised clinical outcome and promotes antibiotic resistance. To address this unmet need, we developed a droplet-in-oil microfluidic device capable of performing antimicrobial susceptibility testing for gonorrhea in <2.5 hours using resazurin-based assay. Our approach is comprised of four sequential processes. First, lyophilization of antimicrobial on microfluidic chip is prepared for future testing. Second, sample containing pathogens is introduced on chip to reconstitute lyophilized reagents. Third, oil is injected to partition sample into individual bacterial incubation reactions. Lastly, bacterial growth under different antimicrobial conditions is examined using resazurin-based assay. We used one ATCC reference strain and two gonorrhea-resistant isolates to demonstrate the accelerated AST capability of our platform.