Evaluation of the Roche LightCycler parvovirus B19 quantification kit for the diagnosis of parvovirus B19 infections

Establishment of functional B cell memory against parvovirus B19 capsid proteins may be associated with resolution of persistent infection

“…Initial serological testing (T ¼ 0), using a B19 VP2 capsid-based immunoassay, for B19 IgG/M proved negative, however B19 DNA positivity was confirmed by both real-time PCR and by dot blot hybridization [Hicks et al, 1995;Braham et al, 2004] (Fig. 1).…”

“…B19 IgG reactivity against linearized VP1 (VP1-D), VP2 (VP2-D), and NS1 was determined using immunoassay formats as described previously [Corcoran et al, 2000]. B19 DNA levels were assessed as described previously [Hicks et al, 1995;Braham et al, 2004] and were quantified using a B19 DNA International Standard preparation [Saldanha et al, 2002]. In addition, cell proliferation studies and cytokine quantitation were carried out as described elsewhere [Corcoran et al, 2000].…”

Identifying the etiologic role of Parvovirus B19 in non-immune hydrops fetalis by histopathology, immunohistochemistry and nucleic acid testing: a retrospective study

“…In this study, a standard immunohistochemistry method with commercially acquired monoclonal antibodies against viral capsid antigens, a commonly-used in-house nested PCR that targets viral capsid regions, and a commercial quantitative Real-time PCR system reported to have a good diagnostic performance were used to detect Parvovirus B19 infection [13,17]. The use of paraffine-embedded tissues for molecular studies may be hindered by the effects of fixatives on target nucleic acids.…”

“…Quantitative detection of viral DNA in purified nucleic acids was performed using the LightCycler Parvovirus B19 Quantification Kit (Roche Diagnostics, Germany) according to the manufacturers' instructions using a LightCycler 2.0 instrument (Roche Diagnostics, Germany) [17].…”

Detection and quantitation of parvovirus B19

“…More recently, both the FDA in America, and the European Pharmocopoeia have determined maximum levels of B19 DNA that are acceptable in plasma pools and their products (Nubling et al, 2004;Tabor and Epstein, 2002). To comply with these regulations and the realization of the value in determining B19 viral titers a number of in-house and commercial assays have been developed to determine B19 levels either by quantitative real-time PCR (Taqman or Light Cycler) or using a form of competitive PCR, as described in this journal (Braham et al, 2004). In addition to their value in screening blood and blood products, these quantitative assays may allow recognition of what degree of viremia is associated with clinical disease, to determine if intervention with immunoglobulin is warranted, and to follow the response to treatment.…”