Detection of a single biomolecule, ranging from nucleic acids, proteins, viruses to bacteria, is of paramount importance in various fields including biology, environment, food and agriculture industry, public health, and medicine. With the understanding of the biological functions of these biomolecules (or bioparticles) and their impacts on public health, environmental pollution, and food safety, advanced detection techniques are unprecedentedly demanded for their early and/or sensitive detection. In this critical review, a series of elegant research about digital detection of biomolecules with potential single copy sensitivity is reviewed and summarized with the focus on the design principle and the innovation of how to accomplish the “digital” detection concept. Starting with a brief introduction on the importance of digital detection, recent advances in “digital” biomolecule detection with single copy sensitivity are grouped and discussed based on the difference of signal reporting systems, including surrogate signal development for “digital” detection, direct visualization for “digital” detection, and nucleic acid amplification enabled “digital” detection. Interdisciplinary combination and integration of different cutting-edge techniques are also discussed with details. The review is closed with the conclusion and future trends.

检测单个生物分子，从核酸、蛋白质、病毒到细菌，在生物学、环境、食品和农业工业、公共卫生和医学等各个领域都至关重要。随着对这些生物分子（或生物粒子）的生物学功能及其对公众健康、环境污染和食品安全的影响的了解，对它们的早期和/或灵敏检测提出了前所未有的先进检测技术要求。在这篇批判性综述中，回顾和总结了一系列关于具有潜在单拷贝敏感性的生物分子数字检测的优雅研究，重点关注设计原理和如何实现“数字”检测概念的创新。从简单介绍数字检测的重要性开始，基于信号报告系统的差异，对具有单拷贝灵敏度的“数字”生物分子检测的最新进展进行了分组和讨论，包括“数字”检测的替代信号开发、“数字”检测的直接可视化和启用核酸扩增的“数字” ”检测。还详细讨论了不同前沿技术的跨学科结合和融合。审查以结论和未来趋势结束。还详细讨论了不同前沿技术的跨学科结合和融合。审查以结论和未来趋势结束。还详细讨论了不同前沿技术的跨学科结合和融合。审查以结论和未来趋势结束。