Han Do Kyoung, Oh Jeonghyun, Lee Jaehyeon, Cho Yong Gon, Park Ju Seong, Choi Jong Soon, Kim Dal Sik, Kwon Joseph
Research Center for Materials Analysis, Korea Basic Science Institute, 169-148, Gwahak-ro, Yuseong-Gu, Daejeon, 34133, Republic of Korea.
Research Center for Materials Analysis, Korea Basic Science Institute, 169-148, Gwahak-ro, Yuseong-Gu, Daejeon, 34133, Republic of Korea; Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, 34134, Republic of Korea.
Biosens Bioelectron. 2021 Mar 15;176:112894. doi: 10.1016/j.bios.2020.112894. Epub 2020 Dec 15.
We report a new paper-based multiplex analytical device (mPAD) for simultaneous screening of three analytes (glutamate dehydrogenase, toxin A, and toxin B) known as biomarkers for Clostridioides difficile infection (CDI). To overcome the limitation of common rapid assays (e.g. lateral flow immunochromatographic and enzyme immunoassays) in terms of multiplexing, sensitivity, simplicity, and ease-of-use, the mPAD is constructed with a three dimensional (3D) configuration of paper components with a multi-channel design. Multiple fluidic paths developed with wax-patterned paper allow the simultaneous detection of glutamate dehydrogenase, toxin A, and toxin B without any cross-reactivity. The 3D fluidic network on the mPAD facilitates a self-operating test procedure for the mixing and addition of amplification reagents with a one-step sliding operation. The results of the multiplex CDI assay can be easily interpreted by the naked eye within 10 min, and are visually intensified over time resulting in up to 3-fold signal amplification. Our device exhibited remarkable analytical performances for the simultaneous detection of three CDI biomarkers, providing a sensitivity of 97%, specificity of 88%, accuracy of 95%, and limits of detection for glutamate dehydrogenase, toxin A, and toxin B of 0.16 ng mL, 0.09 ng mL, and 0.03 ng mL, respectively. These results indicate the high applicability and feasibility of mPAD for multiplex testing for CDI with the advantages of being simple, sensitive, inexpensive, user-friendly, and equipment-free.
我们报告了一种新型的基于纸的多重分析装置（mPAD），用于同时筛查三种被称为艰难梭菌感染（CDI）生物标志物的分析物（谷氨酸脱氢酶、毒素A和毒素B）。为了克服常见快速检测方法（如侧向流动免疫色谱法和酶免疫分析法）在多重检测、灵敏度、简单性和易用性方面的局限性，mPAD采用了具有多通道设计的纸组件三维（3D）结构构建。通过蜡图案化纸形成的多个流体路径可实现谷氨酸脱氢酶、毒素A和毒素B的同时检测，且无任何交叉反应。mPAD上的3D流体网络通过一步滑动操作促进了扩增试剂混合和添加的自操作测试程序。多重CDI检测结果可在10分钟内用肉眼轻松解读，且随着时间推移信号会在视觉上增强，导致信号放大高达3倍。我们的装置在同时检测三种CDI生物标志物方面表现出卓越的分析性能，灵敏度为97%，特异性为88%，准确度为95%，谷氨酸脱氢酶、毒素A和毒素B的检测限分别为0.16 ng/mL、0.09 ng/mL和0.03 ng/mL。这些结果表明mPAD在CDI多重检测方面具有很高的适用性和可行性，具有简单、灵敏、廉价、用户友好且无需设备的优点。
