Wiederkehr, Rodrigo SergioRodrigo SergioWiederkehrMarchal, ElisabethElisabethMarchalFauvart, MaartenMaartenFauvartForceville, TomasTomasForcevilleTaher, AhmedAhmedTaherSteylaerts, TimTimSteylaertsChoe, YoungJaeYoungJaeChoeDusar, HansHansDusarLenci, SilviaSilviaLenciSiouti, EleniEleniSioutiPotsika, Vassiliki T.Vassiliki T.PotsikaAndreakos, EvangelosEvangelosAndreakosStakenborg, TimTimStakenborg2025-04-012025-04-012025-JUN1387-2176WOS:001454297200002https://imec-publications.be/handle/20.500.12860/45472Multiplex polymerase chain reaction (PCR) tests multiple biomarkers or pathogens that cause overlapping symptoms, making it an essential tool in syndromic testing. To achieve a multiplex PCR on chip, a design based on capillary-driven fluidic actuation is proposed. Our silicon chip features 22 reaction chambers and allows primers and probes to be pre-spotted in the reaction chambers prior to use. The design facilitates rapid sample loading through a common inlet channel, delivering reagents to all reaction chambers in less than 10 s. A custom clamping mechanism combined with a double depth cavity design ensures proper sealing during temperature cycling without the need for extra reagents like oil. Temperature cycling and fluorescence imaging were performed using custom-made hardware. As a proof of concept, two single nucleotide polymorphisms (SNPs), CyP2C19*2 and PCSK9 were detected. These results demonstrate the feasibility of on-chip multiplex PCR, compatible with different assays in parallel and requiring only a single pipetting step for reagent loading, without active fluidic actuation like pumping.Journal article10.1007/s10544-025-00745-2WOS:001454297200002REAL-TIME PCRSYSTEMCHIPMEDLINE:40140106