单分子检测是指在单分子水平上通过生物分子的构象变化、动力学、分子之间相互作用以及对单个分子进行操纵等方式进行检测.作为一种新型超灵敏检测手段,单分子检测在生物分子的定量检测领域有广阔的应用前景,尤其单分子荧光检测技术,近年来取得了很大进展,被广泛用于研究多种生物体系.本文对定量单分子检测的最新研究进展进行了系统综述,主要聚焦于定量单分子检测在生物标志物(包括DNA、酶和miRNA等)的超灵敏检测研究中的应用,及其在研究生物分子结构与动态、生物分子间相互作用、生物分子修饰和活细胞检测等方面的应用,本文也对定量单分子检测的发展方向进行了展望. Monomolecular detection refers to the detection of conformational changes of biomolecules, kinetics, interaction between molecules and the manipulation of a single molecule at the level of a single molecule, etc. As a new type of ultra-sensitive detection means, single molecule detection in biological Molecular quantitative detection has broad application prospects, especially single-molecule fluorescence detection technology, made great progress in recent years, has been widely used in the study of a variety of biological systems.In this paper, the quantitative monomolecular detection of the latest research advances were systematically reviewed , Mainly focused on the application of quantitative single-molecule detection in the ultra-sensitive detection of biomarkers (including DNA, enzymes and miRNAs, etc.), and its applications in studying the structure and dynamics of biomolecules, the interactions between biomolecules, the modification of biomolecules Living cell detection and other aspects of the application, this article also quantitative monomolecular detection of the development prospects.