On-Line Dual-Active Valves Based Centrifugal Microfluidic Chip for Fully Automated Point-of-Care Immunoassay

Thereremains an unmet need for a fully integrated microfluidicplatform that can automatically perform multistep and multireagentimmunoassays. Here, we proposed a novel online dual-active valve-basedcentrifugal microfluidic chip, termed DAVM, for fully automatic point-of-careimmunoassay. Practically, the puncture valve, one of the dual activevalves, is capable of achieving precise, on-demand, sequential releaseof prestored reagents, while the other valve-reversible active valveenables controlled retention and drainage of the reaction solutions.Thereby, our technology mitigates the challenges of hydrophilic/hydrophobicmodifications and unstable valve control performance commonly observedin passive valve controls. As a proof of concept, the indirect enzymaticimmunoblotting technique was employed on DAVM for fully automatedimmunological analysis of eight targets, yielding outcomes withinan hour. Furthermore, we conducted a comparative analysis of 28 clinicalsamples with autoimmune diseases. According to 224 clinical data,the sample testing concordance rate between DAVM and the traditionalinstrument was 82%, with a target compliance rate of 97%. Therefore,our DAVM system has powerful potential for fully automated immunoassays.