Aquilina Matthew, Dunn Katherine E
Institute for Bioengineering, University of Edinburgh, Edinburgh, UK.
Deanery of Molecular, Genetic and Population Health Sciences, University of Edinburgh, Edinburgh, UK.
Methods Mol Biol. 2025;2901:203-226. doi: 10.1007/978-1-0716-4394-5_16.
Most biomarker assays are typically designed to detect a single molecule type (DNA, RNA, proteins, etc.) in a single assay. This means that monitoring a diverse biomarker panel could quickly become a complex endeavor, requiring different techniques, labs, and expertise. In this chapter, we describe a method for multiplexed biomarker detection from a single sample using variable-length DNA payload chains as the output signal. Through the course of the assay, payloads are systematically disassembled in the presence of specific biomarkers. The resulting distinctly sized fragments then yield characteristic gel electrophoresis band patterns, which can be detected and quantified using image analysis algorithms. We detail the entire process for constructing DNA payloads and conducting a biomarker detection assay, including the sequence design, laboratory assembly, running the assay, and final image analysis.
大多数生物标志物检测方法通常设计用于在单次检测中检测单一分子类型（DNA、RNA、蛋白质等）。这意味着监测多样化的生物标志物组合可能很快会变得复杂，需要不同的技术、实验室和专业知识。在本章中，我们描述了一种使用可变长度DNA负载链作为输出信号从单个样本中进行多重生物标志物检测的方法。在检测过程中，负载在特定生物标志物存在的情况下会被系统地拆解。产生的大小不同的片段随后会产生特征性的凝胶电泳条带模式，可使用图像分析算法进行检测和定量。我们详细介绍了构建DNA负载和进行生物标志物检测的整个过程，包括序列设计、实验室组装、进行检测以及最后的图像分析。
