Abstract: Microfluidic technology, leveraging its advantages of high throughput, low cost, and rapid detection, is a key technology for nucleic acid point-of-care testing (PoCT). This review systematically summarizes the research progress in microfluidics-based PoCT technology for nucleic acid detection, focusing on nucleic acid purification, nucleic acid amplification, and fluorescence-based nucleic acid detection. Furthermore, the future development directions and challenges are discussed, providing a technical reference for the research and development of PoCT nucleic acid detection devices and instruments.