The present paper aims to prepare a national reference for detection reagents for Yersinia pestis F1 antibody, which was consisted of six positive references, eight negative references, one minimum detection limit reference and one repetitive reference. The uniformity and stability of this reference met the requirements for national references. The collaborative calibration results of four laboratories showed that six positive references were 100% positive, eight negative references were 100% negative, the detection level of the minimum detection limit reference was 1-100 times dilution, and the testing results of repetitive reference were consistent (for the detection reagents based on enzyme-linked immunosorbent assay and up-converting phosphor technology, the coefficient of variation of testing results was less than 15%). So a national reference for detection reagents for Yersinia pestis F1 antibody was prepared which was suitable for the quality evaluation and control of related reagents.