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Diagnosis of erythrovirus B19 relies on serology and the detection of viral DNA. These techniques were believed to detect all field isolates because erythrovirus B19 has been known to undergo little genetic variation (1-2%). Recently, a distinct erythrovirus isolate termed V9, markedly different from erythrovirus B19 (>11% nucleotide disparity), was isolated from a child in France suffering from transient aplastic anemia. Standard PCR assays and serological tests failed to demonstrate an acute erythrovirus B19 infection. Subsequent sequencing of the erythrovirus V9 genome shows that the nucleotide discrepancies encompass the entire genome, indicating that standard erythrovirus B19 PCR assays will not reliably detect erythrovirus V9 DNA. As a tool for studying the epidemiological role and medical importance of this erythrovirus variant, a PCR assay is described that allows simultaneous detection of, and distinction between, erythrovirus B19 and the V9 isolate. Examination of 100 erythrovirus B19 IgM positive samples as well as plasma pools representing 100,000 Danish blood donor units for the presence of B19 and V9 DNA was performed. Despite the apparent absence of erythrovirus V9 in the clinical samples at present, the DNA sequence variability demonstrates that the erythrovirus group may be more divergent than thought previously and the child harboring this isolate may herald erythrovirus V9 as a possible emerging virus., (Copyright 2001 Wiley-Liss, Inc.)

In the B19 VP2 molecule an immunodominant heptapeptide epitope has been detected, recently for which IgG antibodies are synthesized exclusively in the acute phase of B19 infection. Using this acute-phase-specific epitope (KYVTGIN) a 2(nd)-generation epitope-type EIA was developed, which compares serum IgG activity for native VP2 capsids exhibiting conformational VP2 epitopes with IgG activity for the KYVTGIN epitope. In this study the diagnostic performance (clinical sensitivity and specificity) of the 1st and 2nd-generation epitope-type EIAs and of a peptide-based EIA utilising as antigen the KYVTGIN epitope alone was assessed in comparison with various high-quality IgM- and IgG- based B19 assays. Serum samples from 489 patients with B19-related symptoms and asymptomatic controls from three countries were studied. Among 323 patients with B19-IgG, 20% were diagnosed as acute infection, 73% had past immunity and 7% were not classified due to contradictory results among the different assays. The unclassified samples were explored for viral strain diversity by PCR and DNA sequencing but all sequences obtained were B19-like with variance of only a few per cent. The 2nd-generation epitope-type EIA had a diagnostic sensitivity of 98% and a diagnostic specificity of 94%. In combination with conventional approaches, the epitope-type assays increase greatly the accuracy of B19 serodiagnosis., (Copyright 2001 Wiley-Liss, Inc.)