Parvovirus B19 (primate erythroparvovirus 1 or B19V) is a single stranded DNA virus of the genus erythroparvoviridae. It is a respiratory virus with erythroid precursor cell tropism, presenting frequently in children as “fifth disease of childhood”. The majority of infections in adults are asymptomatic or result in non-specific flu-like symptoms. B19V infection can result in complications in immunocompromised and vulnerable individuals, including pregnant women. Transmission of B19V through blood transfusion is well documented from donations containing a high B19V titre, and a small number of probable cases of transfusion-transmitted B19V have been reported in Australia. Three genotypes of B19V are recognised, with genotype 1 responsible for the majority of infections worldwide. The genotypes circulating in Australia are unknown. We aimed to determine the prevalence of B19V DNA among Australian blood donors, as well as quantify the levels of B19V DNA and investigate genotype/s in B19V DNA positive samples. Plasma samples from 4232 donors were collected nationally, and screened in duplicate for the presence of B19V DNA by real-time polymerase chain reaction (PCR). Samples reactive on at least one replicate were tested with a second PCR at an external reference laboratory. Viral load was determined for positive samples by quantitative PCR assay with reference to the WHO B19V standard. Twelve samples tested initially reactive B19V DNA, of which 10 were confirmed (0.23%). Viral loads in the B19V DNA positive samples varied from log 1.90 IU/mL to log 6.25 IU/mL, with the mean B19V DNA level log 3.26 IU/mL (1.58 x 103 IU/mL). Two donations (0.047%) demonstrated a viral load of over 105IU/mL, evidence of in 1 in 2116 donations with potential for transfusion-transmission. Next-generation sequencing is currently being undertaken to determine the B19V genotype/s found in Australian blood donors. As the most common B19V genotype worldwide is genotype 1, it is probable that this is also the predominant genotype circulating in Australia.