An all-in-one microfluidic system via data-driven design for on-site genotyping of genetically modified foods

Divergent global regulations on genetically modified (GM) foods, along with the rise of unauthorized imports and risks of supply-chain contamination, have created an urgent need for rapid, decentralized genotyping tools to ensure food safety and compliance. The development of point-of-care testing has transitioned from centralized polymerase chain reaction (PCR)-based analysis to isothermal amplification platforms that reduce equipment dependency. However, existing systems still require extensive manual handling and external instruments, hindering their field deployment. To address these limitations, we present an all-in-one microfluidic platform that fully integrates sample preparation, recombinase polymerase amplification (RPA), and multiplexed signal readout into a 40 min sample-to-answer workflow. The platform utilizes a novel hybrid active-passive strategy to automate sequential liquid delivery. By employing a data-driven design approach, we engineered specific resistance channels that function as discretized pressure barriers, enabling active, pneumatically driven cascading manipulation of fluid flow for sample lysis. Subsequently, multiplexed RPA analysis can be conducted via passive gravity-driven liquid metering and transfer. Coupled with a docking station containing heating and optics modules and a minipump, the system successfully achieved genotyping of four globally prevalent GM foods (MON 810, Bt 11, NK 603 maize; GTS 40-3-2 soybean), demonstrating excellent sensitivity and specificity with a limit of detection of 0.1% GM content in standard food matrices. This GM food genotyping microfluidic platform paves the way for widespread deployment in customs inspections or field testing, enhances global food safety monitoring, mitigates trade disputes, and supports sustainable agricultural practices.