传统的多重实时荧光聚合酶链式反应(multiplex real-time PCR)通过在一个反应体系中同时加入两对或两对以上的特异性引物，实现对多个靶标的同时检测，但传统定量PCR的多重检测能力有限。将水凝胶与特异性引物结合，构建多重PCR载体，不仅可固定不同靶标的多重引物，还能借助水凝胶编码技术、水凝胶阵列芯片或水凝胶图案化技术，区分结合不同引物的水凝胶，进而实现多重样本的实时快速检测。水凝胶具有高含水量、良好的生物相容性及可调控的三维交联网络等特性，其孔径可通过单体组成与交联方式精确设计，能在维持水相反应的同时提供适度空间约束。调控水凝胶孔隙可提升PCR效率，通过物理或化学固定则可制备水凝胶的阵列芯片。本文简要概述了传统PCR与多重PCR的原理和发展历程，详细阐述了水凝胶多重PCR的原理、优势及研究进展，并对水凝胶PCR检测平台的应用前景及面临的挑战进行了展望。

Traditional multiplex real-time polymerase chain reaction (PCR) enables the simultaneous detection of multiple targets by incorporating two or more pairs of specific primers into a single reaction system. However, traditional quantitative PCR remains limited in its multiplexing capability. By utilizing hydrogels combined with specific primers as a carrier for multiplex PCR, it is possible to immobilize multiple primers targeting different sequences. Furthermore, hydrogels binding distinct primers can be differentiated via hydrogel encoding, array chips, or patterning technologies, thereby facilitating real-time, rapid detection of multiple samples. Characterized by water content, good biocompatibility, and tunable three-dimensional crosslinking networks, hydrogels allow for precise control over size through the adjustment of monomer composition and crosslinking methods. This structural design provides appropriate spatial confinement while maintaining aqueous-phase reaction conditions. Notably, regulating the pore structure of hydrogels enhances PCR efficiency, while physical or chemical fixation enables the fabrication of hydrogel array chips. This paper briefly outlines the principles and developmental history of traditional and multiplex PCR, and comprehensively elaborates on the mechanisms, advantages, and research progress of hydrogel-based multiplex PCR. Finally, the prospects and current challenges of hydrogel PCR detection platforms are discussed.