Abstract: During the COVID-19 pandemic, nucleic acid and its detection technologies became widely recognized among the general population, driven by the need for effective prevention and control measures. This period saw rapid advancements in nucleic acid testing technologies and products, boosting their quality and development. Consequently, there has been an increased focus on improving nucleic acid metrology technologies and the development of certified reference materials, with researchers setting new benchmarks. This review summarizes the current principles, features, and prospective applications of qualitative and quantitative nucleic acid measurement technologies. It highlights the existing shortcomings and potential for growth in nucleic acid metrology research. Moreover, the article systematically reviews the development of nucleic acid certified reference materials, analyzing the varieties, valuation techniques, and field application demands. Finally, the review discusses the emerging opportunities and challenges facing the evolution of nucleic acid metrology technology and certified reference materials.