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Immunodetection of parvovirus B19 virus in human plasma: appraisal of a novel fluorometric immunoassay and comparison with an enzyme immunoassay

Abstract number: 902_p1242

Loftus F.

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Objective:

Human parvovirus B19 is the causative agent for erythema infectiosum or ‘fifth disease’ of children. Symptoms are usually mild in healthy individuals; however, symptoms can be very severe in pregnant women and in the immunocompromised. The aim of the present work is to investigate the performance characteristics of two new viral detection systems, an antigen-capture enzyme immunoassay (EIA) method to detect parvovirus B19 in human plasma and an alternative fluorescence labelling technique (FIA) for enhanced assay sensitivity.

Method:

Recombinant parvovirus B19 capsid protein was used to immunise rabbits and sheep. The antibodies produced were employed in an antigen-capture enzyme immunoassay (EIA) and a fluoroimmunoassay (FIA). The EIA utilised a conventional peroxidase-labelled conjugate, while the FIA utilised a B-phycoerythrin (fluorophore) labelled conjugate. The sensitivity of the assays were evaluated using both purified recombinant parvovirus B19 particles and plasma samples of known parvovirus viraemic load as determined using a validated PCR method.

Results:

Here we show that the sensitivity of the peroxidase labelled EIA is of the order of 5 × 10-7 parvovirus B19 genome equivalents per millilitre. The EIA was able to detect virus in the presence of IgG and IgM antibodies and no false-positive results were obtained. Also we show that the EIA and FIA methods are similar in terms of sensitivity when directed toward the detection of purified recombinant parvovirus capsid protein (VP2).

Conclusion:

EIA and FIA methods as presented are broadly equivalent in terms of specificity towards the parvovirus B19 antigen VP2 capsid protein. Both systems show potential utility in the screening of blood products for the presence of parvovirus B19 antigen.

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Session Details

Date: 01/08/2007
Time: 00:00-00:00
Session name: XXIst ISTH Congress
Subject:
Location: Oxford, UK
Presentation type:
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