Microfluidic-Based Real-Time PCR Platform for Rapid Multiplex Detection of Respiratory Viruses

Rapid and accurate identification of respiratory viruses is essential for effective disease surveillance and clinical management, particularly in settings with limited resources or small sample volumes. In this study, we evaluated the analytical and clinical performance of Genoplex, a microfluidic-based multiplex real-time polymerase chain reaction (PCR) platform currently under development. The Genoplex system enables the simultaneous detection of up to 12 respiratory virus targets within 40 min using a single reaction module. Analytical evaluation demonstrated reliable analytical sensitivity and specificity, with limit of detection ranging from 5 to 500 copies/µL, along with accurate target-specific detection, broad inclusivity across clinically relevant viral strains, and no cross-reactivity or signal interference. Clinical performance was assessed using a large set of 660 nasopharyngeal swab specimens previously tested via routine clinical diagnostic assays. The assay showed high negative percent agreement (99.0–100%) and high positive percent agreement for most targets, with lower agreement observed for adenovirus (72.5%) and human rhinovirus/enterovirus (82.5%). Cohen’s kappa values (κ > 0.8) indicated almost perfect agreement between the Genoplex assay and routine clinical testing. Overall, this study provides an early-stage analytical and clinical evaluation of a developing microfluidic multiplex PCR platform, highlighting its performance characteristics and practical limitations for clinical application.