A capillary-driven microfluidic device for performing spatial multiplex PCR

doi:10.1007/s10544-025-00745-2

A capillary-driven microfluidic device for performing spatial multiplex PCR

2025-06-01

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Rodrigo Sérgio Wiederkehr *

DOI：10.1007/s10544-025-00745-2

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Multiplex 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.

Keyword:

Capillary flow

Multiplex PCR

Silicon chip

Lab-on-a-chip

Fluorescence detection

A capillary-driven microfluidic device for performing spatial multiplex PCR

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