Abstract: Nucleic acid is the carrier of genetic information. With the rapid development of in vitro diagnostic technology based on nucleic acid amplification, nucleic acid detection is widely used in medical diagnosis, forensic identification, import and export inspection and quarantine, species evolution research, etc. Nucleic acid reference materials are a gold standard to ensure the accuracy and traceability of nucleic acid test results. Both the WHO and the Chinese Food and Drug Administration stipulate that in vitro diagnostic reagents need to use international and national reference materials or factory-level standards to guarantee the traceability and accuracy. The analysis of the carcinogenic mechanism of HER2, BRAF, EGFR and other gene mutations has prompted the listing of a series of targeted therapies such as trastuzumab, veilofenib, and rituximab, which has caused a large gap in Chinese nucleic acid reference materials. Due to the complex structure, easy degradation and large molecular weight of nucleic acid, it is difficult to meet the requirements of uniformity and stability. Therefore, development of nucleic acid reference materials is slow. This article briefly reviews preparation methods of nucleic acid reference materials and domestic and abroad research progress, for the efficient development of nucleic acid reference materials for in vitro diagnostics.