Recent advances in phenotypic antimicrobial susceptibility testing enabled by microfluidic technologies

Abstract

Antimicrobial resistance (AMR) poses an urgent global health threat, driving the need for rapid and accurate antimicrobial susceptibility testing (AST). Traditional phenotypic AST methods remain the clinical gold standard but are hindered by prolonged turnaround times and labor-intensive procedures. Microfluidic technologies have emerged as transformative platforms, enabling miniaturized, high-throughput, and integrated phenotypic AST workflows with accelerated result delivery. This review comprehensively summarizes recent advances in microfluidic phenotypic AST, categorizing platforms by cultivation strategies—such as static chambers, flow chambers, SlipChip variants, and hybrid droplet-chamber systems—and surveying diverse signal detection modalities including fluorescence, label-free imaging, Raman, electrical, and mechanical readouts, each offering distinct advantages and limitations. Key innovations such as concentration gradient generation, digital single-cell manipulation, and AI-enhanced image analysis have significantly improved sensitivity, speed, and clinical applicability. However, widespread adoption remains challenged by sample-to-result integration, slow-growing pathogens, interference from residual antibiotics, and the lack of robust standardization. We further discuss emerging solutions, including automated sample preparation, multimodal detection, and computational data fusion, and outline future opportunities for translating microfluidic phenotypic AST into routine diagnostics. Collectively, these advances hold substantial promise for combating AMR by enabling personalized, rapid, and actionable antimicrobial therapy.