Simple manipulation of enzyme-linked immunosorbent assay (ELISA) using an automated microfluidic interface.
Simple manipulation of enzyme-linked immunosorbent assay (ELISA) using an automated microfluidic interface.
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DOI：
10.1039/d2ay00326k
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发表时间：
2022-05-13
影响因子：
3.1
通讯作者：
Henry, Charles S.
中科院分区：
文献类型：
--
作者：
Panraksa, Yosita;Jang, Ilhoon;Carrell, Cody S.;Amin, Anita G.;Chailapakul, Orawon;Chatterjee, Delphi;Henry, Charles S.
在侧流免疫分析 (LFIA) 平台中，基于酶的 LFIA 提供信号放大功能以提高灵敏度。然而，大多数基于酶的 LFIA 需要多个定时步骤，这使得它们在即时检测 (POCT) 中的实用性变得复杂。在这里，我们报告了一种用于 LFIA 的微流体接口，可自动添加样品、缓冲液和试剂，大大简化操作，同时实现与更复杂的测定相关的高分析严格性。微流体接口还保持了标准 LFIA 的低成本和小占地面积。该平台由聚酯薄膜、双面胶带和硝化纤维组合而成，适合手掌大小。所有试剂都在硝化纤维上干燥，以方便顺序试剂输送，并且样品用作洗涤缓冲液以最大程度地减少步骤。添加样品后，用户只需等待 15 分钟即可获得比色结果。本手稿讨论了通道几何形状的开发和优化，以实现简单的步骤酶放大免疫测定。作为概念验证目标，我们选择了阿拉伯脂甘露聚糖 (LAM)，这是一种世界卫生组织确定的活动性结核病的尿液生物标志物，以证明该设备的可行性和可靠性。结果显示，该设备在样品加载后 15 分钟内成功检测到磷酸盐缓冲液 (PBS) 以及加标尿样中的 LAM。颜色变化的最小浓度为 25 ng/mL。
Among lateral flow immunoassay (LFIA) platforms, enzyme-based LFIAs provide signal amplification to improve sensitivity. However, most enzyme-based LFIAs require multiple timed steps, complicating their utility in point-of-care testing (POCT). Here, we report a microfluidic interface for LFIAs that automates sample, buffer, and reagent addition, greatly simplifying operation while achieving the high analytical stringency associated with more complex assays. The microfluidic interface also maintains the low cost and small footprint of standard LFIAs. The platform is fabricated from a combination of polyester film, double-sided adhesive tape, and nitrocellulose, and fits in the palm of your hand. All reagents are dried on the nitrocellulose to facilitate sequential reagent delivery, and the sample is used as the wash buffer to minimize steps. After the sample addition, a user simply waits 15 min for a colorimetric result. This manuscript discusses the development and optimization of the channel geometry to achieve a simple step enzyme amplified immunoassay. As a proof-of-concept target, we selected lipoarabinomannan (LAM), a WHO identified urinary biomarker of active tuberculosis, to demonstrate the device feasibility and reliability. The results revealed that the device successfully detected LAM in phosphate buffer (PBS) as well as spiked urine samples within 15 min after sample loading. The minimum concentration of color change was achieved at 25 ng/mL.
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DOI：
10.1172/jci140461
发表时间：
2020-11-02
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Broger T;Nicol MP;Sigal GB;Gotuzzo E;Zimmer AJ;Surtie S;Caceres-Nakiche T;Mantsoki A;Reipold EI;Székely R;Tsionsky M;van Heerden J;Plisova T;Chikamatsu K;Lowary TL;Pinter A;Mitarai S;Moreau E;Schumacher SG;Denkinger CM
通讯作者：
Denkinger CM
影响因子：
8.4
作者：
Jang, Ilhoon;Berg, Kathleen E.;Henry, Charles S.
通讯作者：
Henry, Charles S.
影响因子：
8.9
作者：
Jang, Ilhoon;Carrao, Daniel B.;Henry, Charles S.
通讯作者：
Henry, Charles S.
影响因子：
7.4
作者：
Channon, Robert B.;Nguyen, Michael P.;Dandy, David S.
通讯作者：
Dandy, David S.
影响因子：
3.9
作者：
Brogan, KL;Shin, JH;Schoenfisch, MH
通讯作者：
Schoenfisch, MH