Fully automated centrifugal microfluidic system for self-calibrating isothermal nucleic acid quantification

We present a fully automated centrifugal microfluidic system to enable self-calibrating isothermal nucleic acid quantification. The disc comprises two modules: one generates an on-disc calibration curve through the amplification of serially diluted standard samples, and the other performs cell lysis, nucleic acid extraction, and isothermal amplification of loaded target samples to yield quantifiable signals. This integrated design allows accurate and consistent calibration-based quantification under shared on-disc reaction conditions for both standards and target samples, providing reliable analysis even under variable testing conditions. The system successfully quantified Escherichia coli O157:H7 in minced rump samples spiked with the target pathogen and then pre-enriched, and the on-disc analysis was completed within 1 h after sample loading without further manual intervention. The concurrent generation of a standard curve on the same chip enhances robustness against reaction variability, positioning the platform as a practical tool for future molecular diagnostics. This approach could facilitate more reliable quantitative molecular diagnostics in decentralized testing environments.