Background: Parvovirus B19 (PVB19) is transmitted via transfusion of blood and blood products. PVB19 is resistant to viral inactivation methods, which poses a threat to blood safety. We investigated the prevalence of PVB19 antibodies and DNA in healthy blood donors from the South African National Blood Bank Service to evaluate the necessity of PVB19 DNA testing. Study Design and Method: A retrospective analysis of 1500 residual plasma specimens from healthy blood donors from the SANBS repository were screened in mini- pools of 20 for PVB19 DNA using a quantitative polymerase chain reaction (PCR). Positive pools were resolved by individual viral load testing and screened for PVB19 immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies to correlate viral loads with serological status. PVB19 IgG prevalence was determined by testing 90 randomly selected specimens from the 1500 plasma specimens. Results: The prevalence of PVB19 IgG, IgM and IgG, and DNA was 62.2%, 0.06%, and 0.9%, respectively. Fourteen of the 1500 blood donor specimens received, had detectable PVB19 viral loads. Nine of the fourteen donors with detectable viral loads were PVB19 IgG seropositive. The PVB19 viral loads ranged from 1.81 to 5.32 log IU/mL. Four of the fourteen viraemic donors had a viraemia > 10(4) IU/mL. Conclusion: We have demonstrated a low prevalence of PVB19 DNA in SANBS blood donors. The predominance of low-level viraemia and the presence of PVB19 antibodies, suggests that the risk of transfusion transmission of PVB19 among SANBS donors may be relatively low.