Mandatory requirements for plasma pools containing a maximum concentration of human parvovirus B19 DNA will be established in Taiwan in the near future. To facilitate the implementation of the policy, a National Standard and working reagent were established for human parvovirus B19 DNA nucleic acid amplification techniques (NAT). The standards are intended to be used for the quality control of B19 NAT assays and as a quantitative reference material for B19 NAT testing. A collaborative study, including 10 laboratories from seven countries, was done in order to establish the National Standard and working reagent for B19 DNA NAT assays by calibration, in International Units (IU), against the WHO International Standard for B19 (99 /800). Participants were requested to test the candidate materials and the WHO International Standard by the NAT assay in routine use in the laboratory. The mean B19 DNA content of the standards was determined by each laboratory in three independent assays and the results were analyzed statistically. Overall, a high level of agreement among results was achieved from different laboratories. Consequently, the first National Standard and working reagent for B19 DNA NAT assays with an assigned value of 1.9 × 106 IU/mL and 2.0 × 104 IU/ mL, respectively, were established. The results of the stability analysis indicated that both reagents were stable for 4 weeks at 25°C, for 8 weeks at 4°C, for at least 18 months at -20°C or -80°C.