An integrated IoT microfluidic platform for vector-borne pathogen detection via closed-loop valve control and...

Vector-borne diseases, including arboviral infections and malaria, pose a major global health threat and underscore the need for rapid, accurate, and multiplex nucleic acid detection technologies for clinical diagnosis and surveillance. However, previous studies on reagent-preloaded and switching valve-based microfluidic cartridge platforms have insufficiently addressed several key engineering challenges, particularly the reliable control of multichannel switching valves and the inherently critical requirement for uniform reagent distribution across multiple reaction zones. These limitations affect system stability and repeatability, thereby constraining their practical applicability in RNA-based arbovirus detection and DNA-based Plasmodium genotyping. We developed a Microfluidic Vector-borne Pathogen Detection (MVPD) system that introduced comprehensive advancements in fluidic architecture, reagent management, and system informatization. The MVPD system utilized a six-reaction tube microfluidic cartridge that integrated a multichannel switching valve actuated by a magnetic field-based closed-loop control mechanism, along with a dual-mode fluid-handling strategy that combined a peristaltic pump with a constant positive-pressure source for precise fluidic manipulation. Coupled with a magnetic separation module, thermal cycling unit, and six-channel optical detection, the MVPD system achieved independently verified nucleic acid extraction with > 90% efficiency and reliable amplification of up to 36 targets with strong linearity, while completing the entire sample-in-answer-out workflow around 60 min. It further demonstrated high sensitivity, broad sample compatibility, long-term stability, specificity, and reproducible multi-target parallel detection, all in a fully integrated sample-in-answer-out manner. Multi-center testing of 417 port blood samples confirmed 100% concordance with reference methods, while real-time cloud-based data management enabled remote monitoring and efficient cross-regional surveillance. Collectively, these features showed that the MVPD system effectively addressed the critical challenges of multichannel valve control and uniform fluid distribution, providing a robust, fully automated solution for vector-borne pathogen detection.