Automated multi-diagnostic platform for respiratory viruses using integrated microfluidic cartridge and RT-RPA/CRISPR detection

초록

Conventional molecular diagnostics offer high sensitivity and specificity, but lengthy workflows, complex protocols, and laboratory-grade instrumentation limit their point-of-care application. We developed a fully automated sample-to-answer molecular diagnostic platform consisting of a disposable microfluidic cartridge and a compact detection station. By loading a clinical specimen mixed with magnetic beads into the cartridge and operating the station, the system executes a hands-off sequence-including virus capture, washing, thermal lysis, RT-RPA, and CRISPR-Cas12a fluorescence detection-without additional equipment or prior nucleic-acid extraction. The platform simultaneously detects three high-priority respiratory viruses-influenza A (H1N1 and H3N2), influenza B, and SARS-CoV-2-within 35-50 min. The analytical performance was evaluated using synthetic template DNAs, cultured viruses, and clinical specimens. On-cartridge detection limits were 1, 100, 10, and 100 copies center dot mu L-1 for H1N1, H3N2, influenza B, and SARS-CoV-2, respectively. In 30 nasopharyngeal samples, the platform achieved 100% sensitivity and specificity for influenza A and B, and 100% sensitivity with 95.2% specificity for SARS-CoV-2 compared to RT-PCR. These results show that integrated automation facilitates hands-off testing without pre-extraction or additional equipment, providing a practical path to rapid, multiplexed point-of-care diagnosis of respiratory viruses.

키워드