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一种用于多重比色法检测蛋白质生物标志物的便携式低成本离心微流控平台。

A portable and low-cost centrifugal microfluidic platform for multiplexed colorimetric detection of protein biomarkers.

机构信息

KTH Royal Institute of Technology, Division of Nanobiotechnology, Department of Protein Science, Science for Life Laboratory, Solna, Sweden.

KTH Royal Institute of Technology, Division of Nanobiotechnology, Department of Protein Science, Science for Life Laboratory, Solna, Sweden.

出版信息

Anal Chim Acta. 2023 Mar 8;1245:340823. doi: 10.1016/j.aca.2023.340823. Epub 2023 Jan 10.

Abstract

Cytokines play a very important role in our immune system by acting as mediators to put up a coordinated defense against foreign elements in our body. Elevated levels of cytokines in the body can signal to an ongoing response of the immune system to some abnormality. Thus, the quantification of a panel of cytokines can provide valuable information regarding the diagnosis of specific diseases and state of overall health of an individual. Conventional Enzyme Linked Immunosorbent Assay (ELISA) is the gold-standard for quantification of cytokines, however the need for trained personnel and expensive equipment limits its application to centralized laboratories only. In this context, there is a lack of simple, low-cost and portable devices which can allow for quantification of panels of cytokines at point-of-care and/or resource limited settings. Here, we report the development of a versatile, low-cost and portable bead-based centrifugal microfluidic platform allowing for multiplexed detection of cytokines with minimal hands-on time and an integrated colorimetric signal readout without the need for any external equipment. As a model, multiplexed colorimetric quantification of three target cytokines i.e., Tumor necrosis factor alpha (TNF-α), Interferon gamma (IFN-γ) and Interleukin-2 (IL-2) was achieved in less than 30 min with limits of detection in ng/mL range. The developed platform was further evaluated using spiked-in plasma samples to test for matrix interference. The ease of use, low-cost and portability of the developed platform highlight its potential to serve as a sample-to-answer solution for detection of cytokine panels in resource limited settings.

摘要

细胞因子在我们的免疫系统中起着非常重要的作用,作为介质,它们协调一致地抵抗体内的外来物质。体内细胞因子水平的升高表明免疫系统对某些异常情况的持续反应。因此,细胞因子谱的定量可以提供有关特定疾病诊断和个体整体健康状况的有价值的信息。传统的酶联免疫吸附测定 (ELISA) 是细胞因子定量的金标准,但是需要经过培训的人员和昂贵的设备,这限制了它只能在集中式实验室中使用。在这种情况下,缺乏简单、低成本和便携式的设备,无法在床边或资源有限的环境中对细胞因子谱进行定量。在这里,我们报告了一种多功能、低成本和便携式的基于珠子的离心微流控平台的开发,该平台允许在最少的人工操作时间内进行细胞因子的多重检测,并具有集成的比色信号读出功能,而无需任何外部设备。作为模型,我们实现了三种靶细胞因子(TNF-α、IFN-γ 和 IL-2)的多重比色定量,整个过程不到 30 分钟,检测限达到 ng/mL 范围。该开发平台还使用添加的血浆样本进行了评估,以测试基质干扰。该平台易于使用、成本低且便携,突出了其在资源有限的环境中作为用于检测细胞因子谱的即插即用解决方案的潜力。

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