Capillary flow velocity-based length identification of PCR and RPA products on paper microfluidic chips.

IF 10.7 1区 生物学 Q1 BIOPHYSICS Biosensors and Bioelectronics Pub Date : 2024-10-22 DOI:10.1016/j.bios.2024.116861
Bailey C Buchanan, Reid S Loeffler, Rongguang Liang, Jeong-Yeol Yoon
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Abstract

This work demonstrates a novel, non-fluorescence approach to the length identification of polymerase chain reaction (PCR) and recombinase polymerase amplification (RPA) products, utilizing capillary flow velocities on paper microfluidic chips. It required only a blank paper chip, aminated microspheres, and a smartphone, with a rapid assay time and under ambient lighting. A smartphone evaluated the initial capillary flow velocities on the paper chips for the PCR and RPA products from various bacterial samples, where the pre-loaded aminated microspheres differentiated their flow velocities. Flow velocities were analyzed at different time frames and compared with the instantaneous flow velocities and interfacial tension (γLV) data. Subsequent error analysis justified the use of the initial time frames. A robust linear relationship could be established between the initial flow velocities against the square root of the product lengths, with R2 values of 0.981 for PCR and 0.993 for RPA. The assay seemed not to have a significant dependency on the cycle numbers and initial target concentrations. This novel method can be potentially used with various paper microfluidic methods of nucleic acid amplification tests towards rapid and handheld assays.

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基于毛细管流速的纸质微流控芯片 PCR 和 RPA 产物长度鉴定。
这项工作展示了一种利用纸质微流控芯片上的毛细管流速对聚合酶链反应(PCR)和重组酶聚合酶扩增(RPA)产物进行长度鉴定的非荧光新方法。它只需要一个空白纸质芯片、氨基化微球和一部智能手机,在环境光下即可进行快速检测。智能手机评估了纸芯片上来自各种细菌样本的 PCR 和 RPA 产物的初始毛细管流速,其中预装的胺化微球可区分它们的流速。对不同时间段的流速进行了分析,并与瞬时流速和界面张力(γLV)数据进行了比较。随后的误差分析证明使用初始时间框架是正确的。在初始流速与产物长度的平方根之间建立了稳健的线性关系,PCR 的 R2 值为 0.981,RPA 为 0.993。该检测方法似乎与循环次数和初始目标浓度没有明显的关系。这种新方法可与各种纸质微流控核酸扩增检测方法结合使用,实现快速和手持式检测。
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来源期刊
Biosensors and Bioelectronics
Biosensors and Bioelectronics 工程技术-电化学
CiteScore
20.80
自引率
7.10%
发文量
1006
审稿时长
29 days
期刊介绍: Biosensors & Bioelectronics, along with its open access companion journal Biosensors & Bioelectronics: X, is the leading international publication in the field of biosensors and bioelectronics. It covers research, design, development, and application of biosensors, which are analytical devices incorporating biological materials with physicochemical transducers. These devices, including sensors, DNA chips, electronic noses, and lab-on-a-chip, produce digital signals proportional to specific analytes. Examples include immunosensors and enzyme-based biosensors, applied in various fields such as medicine, environmental monitoring, and food industry. The journal also focuses on molecular and supramolecular structures for enhancing device performance.
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