A capillary-driven microfluidic device for performing spatial multiplex PCR

IF 3 4区 医学 Q3 ENGINEERING, BIOMEDICAL
Rodrigo S. Wiederkehr, Elisabeth Marchal, Maarten Fauvart, Tomas Forceville, Ahmed Taher, Tim Steylaerts, YoungJae Choe, Hans Dusar, Silvia Lenci, Eleni Siouti, Vassiliki T. Potsika, Evangelos Andreakos, Tim Stakenborg
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引用次数: 0

Abstract

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.

一种用于执行空间多重PCR的毛细管驱动微流控装置
多重聚合酶链反应(PCR)检测引起重叠症状的多种生物标志物或病原体,使其成为综合征检测的重要工具。为了在芯片上实现多重PCR,提出了一种基于毛细管驱动的流体驱动设计。我们的硅芯片具有22个反应室,并允许在使用前在反应室中预点出引物和探针。该设计便于快速样品加载通过一个共同的入口通道,提供试剂到所有的反应室在不到10秒。定制夹紧机构与双深度腔体设计相结合,确保在温度循环期间适当密封,而无需额外的试剂,如油。温度循环和荧光成像使用定制的硬件。作为概念证明,检测到两个单核苷酸多态性(snp), CyP2C19*2和PCSK9。这些结果证明了片上多重PCR的可行性,它兼容不同的平行测定,只需要一个移液步骤就可以装载试剂,而不需要主动流体驱动,如泵送。
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来源期刊
Biomedical Microdevices
Biomedical Microdevices 工程技术-工程:生物医学
CiteScore
6.90
自引率
3.60%
发文量
32
审稿时长
6 months
期刊介绍: Biomedical Microdevices: BioMEMS and Biomedical Nanotechnology is an interdisciplinary periodical devoted to all aspects of research in the medical diagnostic and therapeutic applications of Micro-Electro-Mechanical Systems (BioMEMS) and nanotechnology for medicine and biology. General subjects of interest include the design, characterization, testing, modeling and clinical validation of microfabricated systems, and their integration on-chip and in larger functional units. The specific interests of the Journal include systems for neural stimulation and recording, bioseparation technologies such as nanofilters and electrophoretic equipment, miniaturized analytic and DNA identification systems, biosensors, and micro/nanotechnologies for cell and tissue research, tissue engineering, cell transplantation, and the controlled release of drugs and biological molecules. Contributions reporting on fundamental and applied investigations of the material science, biochemistry, and physics of biomedical microdevices and nanotechnology are encouraged. A non-exhaustive list of fields of interest includes: nanoparticle synthesis, characterization, and validation of therapeutic or imaging efficacy in animal models; biocompatibility; biochemical modification of microfabricated devices, with reference to non-specific protein adsorption, and the active immobilization and patterning of proteins on micro/nanofabricated surfaces; the dynamics of fluids in micro-and-nano-fabricated channels; the electromechanical and structural response of micro/nanofabricated systems; the interactions of microdevices with cells and tissues, including biocompatibility and biodegradation studies; variations in the characteristics of the systems as a function of the micro/nanofabrication parameters.
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