面向核酸检测的连续流PCR微流控技术研究进展

Advances in Continuous-Flow PCR Microfluidics for Nucleic Acid Detection

摘要: 聚合酶链反应作为核酸检测的核心技术，在生命科学研究与临床诊断中发挥着不可替代的作用。传统热循环仪存在检测周期长、设备体积庞大、试剂消耗量大等局限，难以满足现场快速检测的多元需求。连续流PCR通过固定温区、流体跨区流动的模式实现热循环，规避了温控系统热惯性的影响，兼具扩增速度快、试剂消耗量低、易集成小型化等优势，是微流控核酸检测领域核心研究方向。迄今，已发展逶迤形、三维螺旋、振荡流、闭环循环流、液滴连续流等多类构型。在结构设计、性能优化与场景应用层面均取得了长足进展。本文系统梳理了连续流PCR的技术原理与结构演化脉络，阐述了该技术在临床即时检测、食品安全检测、环境监测等领域的应用现状。

Abstract: Polymerase chain reaction (PCR), as a core technology for nucleic acid detection, plays an indispensable role in life science research and clinical diagnosis. However, conventional thermal cyclers suffer from limitations such as long detection cycles, bulky instrumentation, and high reagent consumption, which hinder their ability to meet the diverse demands of on-site rapid testing. Continuous-flow PCR (CF-PCR) achieves thermal cycling by employing spatially fixed temperature zones with the fluid flowing across them, thereby circumventing the thermal inertia inherent in temperature control systems. This approach offers advantages including fast amplification, low reagent consumption, and ease of integration and miniaturization, establishing it as a key research direction in the field of microfluidic nucleic acid detection. To date, various configurations have been developed, including serpentine channels, three-dimensional helical structures, oscillating-flow systems, closed-loop circulatory designs, and droplet-based continuous-flow formats. Significant progress has been made in structural design, performance optimization, and application scenarios. This review systematically summarizes the working principles and structural evolution of continuous-flow PCR, and presents the current status of its applications in clinical point-of-care testing, food safety detection, and environmental monitoring.