关键词: 数字PCR, 标准物质, 生物标准物质, 测量不确定度

Abstract: In the field of reference material development, the development of biological reference material has gradually become a hot topic. At the same time, the development and application of nucleic acid-based detection technology has promoted the process of nucleic acid reference material. Nucleic acid reference materials require high-level and accurate valuation methods, and digital PCR has been widely used as a single-molecule quantitative technology. Digital PCR is an absolute quantitative method for the determination of nucleic acid molecules. For example, droplet digital PCR uses droplets formed by water-in-oil as the reaction chamber, and distributes the reaction solution containing the DNA template to a large number of independent reaction chambers for expansion. Increase the reaction, and then quantify the copy number of DNA by counting the positive signals in the reaction chamber, so as to achieve the purpose of accurately quantifying the copy number of nucleic acid. This article reviewed the literature on the research of digital PCR and  the latest application progress of digital PCR technology in the development of DNA and nucleic acid reference materials, especially in the fields of medical diagnosis and genetic modification detection, which was expected to provide reference for the research of reference material development.

Key words: digital PCR, reference material, biological reference material, measurement uncertainty