Fengjiao Zhu, Songnan Liu, Xue Bai, Xianming Liu, Bingcheng Lin, Yao Lu.Analytical and Bioanalytical Chemistry 2023, 415 Advances in optical counting and imaging of micro/nano single-entity reactors for biomolecular analysis. Analytical and Bioanalytical Chemistry 2023, 142 Single-molecule fluorescence methods for protein biomarker analysis. Haihan He, Chuhong Wu, Muhammad Saqib, Rui Hao.Tyramine signal amplification on polystyrene microspheres for highly sensitive quantification of Aflatoxin B1 in peanut samples. Rui Chen, Chen Zhan, Chenxi Huang, Qifu He, Junwang Bao, Xiuwen Zhang, Zhixiong Pi, Yiping Chen.One-step detection of T4 polynucleotide kinase activity based on single particle-confined enzyme reaction and digital particle counting. Dailu Jia, Wenjiao Fan, Wei Ren, Chenghui Liu.A microchamber-free and enzyme-free digital assay based on ultrabright fluorescent microspheres. Feng Gong, Yixia Yang, Xiaoyun Shan, Zhiyou Tan, Jingjing Qian, Songbai Tian, Xinghu Ji, Zhike He.Biosensors and Bioelectronics: X 2023, 13, 100312. Droplet-free digital immunoassay based on electrochemiluminescence. Single Molecule Upconversion-Linked Immunosorbent Assay with Extended Dynamic Range for the Sensitive Detection of Diagnostic Biomarkers. Mickert, Antonín Hlaváček, Petr Skládal, and Hans H. Cu/Mn Double-Doped CeO2 Nanocomposites as Signal Tags and Signal Amplifiers for Sensitive Electrochemical Detection of Procalcitonin. Zhe-Han Yang, Shirong Ren, Ying Zhuo, Ruo Yuan, and Ya-Qin Chai.Catalyzed Deposition of Signal Reporter for Highly Sensitive Surface-Enhanced Raman Spectroscopy Immunoassay Based on Tyramine Signal Amplification Strategy. Cuicui Fu, Sila Jin, Wenbing Shi, Joohee Oh, Haiyan Cao, Young Mee Jung.A Single-Molecule Homogeneous Immunoassay by Counting Spatially “Overlapping” Two-Color Quantum Dots with Wide-Field Fluorescence Microscopy. Xiaojun Liu, Conghui Huang, Chenghua Zong, Aiye Liang, Zhangjian Wu, Yusu Zhang, Qingquan Zhang, Wenfeng Zhao, Hongwei Gai.Wash- and Amplification-Free Digital Immunoassay Based on Single-Particle Motion Analysis. Kenji Akama, Niina Iwanaga, Koya Yamawaki, Masaki Okuda, Krupali Jain, Hiroshi Ueno, Naoki Soga, Yoshihiro Minagawa, Hiroyuki Noji.Single Molecule Protein Detection with Attomolar Sensitivity Using Droplet Digital Enzyme-Linked Immunosorbent Assay. Limor Cohen, Naiwen Cui, Yamei Cai, Padric M.Simplified Digital Enzyme-Linked Immunosorbent Assay Using Tyramide Signal Amplification and Fibrin Hydrogels. Microchamber-Free Digital Flow Cytometric Analysis of T4 Polynucleotide Kinase Phosphatase Based on Single-Enzyme-to-Single-Bead Space-Confined Reaction. Lijun Zhang, Wenjiao Fan, Dailu Jia, Qinya Feng, Wei Ren, Chenghui Liu.High-Throughput, High-Multiplex Digital Protein Detection with Attomolar Sensitivity. Bifunctional Pdots-Based Novel ECL Nanoprobe with Qualitative and Quantitative Dual Signal Amplification Characteristics for Trace Cytokine Analysis. Jinxia Liu, Xiaodi Liu, Huanhuan Chen, Luxia Yang, Aiting Cai, Haiwei Ji, Qi Wang, Xiaobo Zhou, Guo Li, Mingmin Wu, Yuling Qin, Li Wu.Future of Digital Assays to Resolve Clinical Heterogeneity of Single Extracellular Vesicles. Digital Protein Detection in Bulk Solutions. ![]() Sungjun Beck, Donghae Shin, Sun Jin Kim, Per Niklas Hedde, Weian Zhao.This article is cited by 68 publications. Our method has potential applications in simple in vitro diagnostic systems for detecting ultralow concentrations of protein biomarkers. ![]() The obtained limit of detection represents a >20-fold higher sensitivity than conventional ELISA. ![]() The performance of our method was demonstrated by detecting hepatitis B surface antigen with a limit of detection of 0.09 mIU/mL (139 aM) and a dynamic range of over 4 orders of magnitude. Consequently, the fluorescence signal is localized on a portion of the beads, making it possible to count the number of labeled beads digitally. By adjusting the bead concentration in the HRP-tyramide reaction and conducting the reaction using freely moving beads, tyramide radicals are deposited only on beads labeled with HRP and there is no diffusion to other beads. In our method, tyramide substrate reacts with horseradish peroxidase (HRP) labeled with an immunocomplex on beads, and the substrate is converted into short-lived radical intermediates. This study presents a novel Digital ELISA that does not require droplets instead, enzyme reaction products are concentrated using a tyramide signal amplification system. The key aspect of this technology is to concentrate enzyme reaction products from a single target molecule in femtoliter droplets. Digital enzyme-linked immunosorbent assay (ELISA) is a single molecule counting technology and is one of the most sensitive immunoassay methods.
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