In order to improve serodiagnostic methods for the determination of the state of human parovirus B19 infection, a new test system, recomLine Parvovirus, based on the use of recombinant antigens, has been developed and evaluated. The test system combines the advantages of enzyme-linked immunosorbent assay (ELISA) methods with those of the Western blot technique. For the recombinant line assay, five antigens of human parvovirus B19 that were recombinantly produced in Escherichia coli were applied directly on nitrocellulose membranes: VP2, the aminoterminal and the carboxyterminal domain of VP1 (VP-N and VP-C), VP-1S another fragment of VP-N and NS1. In addition, empty virus particles isolated from eukaryotic cell cultures were also applied. The recombinant-line assay was used to detect human IgG and IgM antibodies directed against human parvovirus B19. In addition, the avidity of the IgG antibodies was investigated. The recombinant line assay was evaluated using 87 human serum samples of patients recently infected with human parvovirus B19 including 10 samples of three infection time courses and 100 serum samples of healthy blood donors. All results were compared with commercially available ELISAs. In the case of discrepancies, Western blot analysis was performed. The data revealed the recombinant line assay to be highly sensitive and specific. The individual determination of the human immune response against several recombinant antigens covering the structural proteins of human parvovirus B19 gives a deeper insight into the actual status of infection. In addition, the determination of IgG avidity against these individual recombinant antigens enables a more precise and differentiated picture of the infection event.