Detection of rubella virus-specific immunoglobulin M antibodies with a baculovirus-expressed E1 protein

Schmidt, Michel; Lindqvist, Christer; Salmi, A.; Oker‐Blom, Christian

doi:10.1128/cdli.3.2.216-218.1996

Cited by 7 publications

(1 citation statement)

References 40 publications

(52 reference statements)

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“…Most commercial tests incorporate lysates or extracts of T. gondii or R. virus as specific antibody capture molecules. However, there are technical limitations to their production, such as the need to maintain living parasites and difficulties in the standardisation of cultures ( Schmidt et al 1996 , Pfrepper et al 2005 , Holec-Gasior 2013 ). For these reasons, there are several reports of evaluations of recombinant proteins by EIAs that were designed to replace extracts and lysates of T. gondii ( Holec-Gasior 2013 ) and R. virus ( Schmidt et al 1996 , Liu et al 2013 ) used in current tests.…”

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Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella

Baschirotto

Krieger

Foti

2017

Mem. Inst. Oswaldo Cruz

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BACKGROUND During pregnancy, toxoplasmosis and rubella can cause serious damage to the mother and the foetus through vertical transmission. Early diagnosis enables implementation of health measures aimed at preventing vertical transmission and minimising damage caused by these diseases.OBJECTIVE Here, we report the development of a multiplex assay for simultaneous detection of IgG antibodies produced during toxoplasmosis and rubella infection.METHODS This assay is based on xMap technology. Initially, by singleplex assays, we evaluated the following antigens: one Toxoplasma gondii lysate; two antigenic extracts of T. gondii (TOX8131 and TOX8122); fragments of T. gondii antigens [SAG-1 (amino acids 45-198), GRA-7 (24-100), GRA-1 (57-149), ROP-4, and MIC-3 (234-306)]; two chimeric antigens composed of fragments of SAG-1, GRA-7, and P35 (CTOX and CTOXH); and fragments of Rubella virus antigens [E-1 (157-176, 213-239, 374-390), E-2 (31-105), and C (1-123)].FINDINGS A multiplex assay to simultaneously diagnose toxoplasmosis and rubella was designed with the best-performing antigens in singleplex and multiplex assays, which included CTOXH, T. gondii lysate, TOX8131, E-1, and E-2. The multiplex assay showed 100% sensitivity and specificity for anti-T. gondii IgG detection and 95.6% sensitivity and 100% specificity for anti-R. virus IgG detection.MAIN CONCLUSIONS We found that, despite the difficulties related to developing multiplex systems, different types of antigens (extracts and recombinant proteins) can be used to develop high-performance diagnostic tests. The assay developed is suitable to screen for prior T. gondii and R. virus infections, because it is a rapid, high-throughput, low-cost alternative to the current standard diagnostic tools, which require multiple individual tests.

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Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella

Baschirotto

Krieger

Foti

2017

Mem. Inst. Oswaldo Cruz

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Baculovirus-Mediated Large-Scale Expression and Purification of a Polyhistidine-Tagged Rubella Virus Capsid Protein

Schmidt

Tuominen

Johansson

et al. 1998

Protein Expression and Purification

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Preparation of immunoblot test stripes from a Rubella virus-like particles dye crystal complex as antigen

et al. 2005

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Stably transfected Chinese hamster ovary (CHO24S) cells were the source for Rubella virus-like particles (RVLP) containing all structural proteins (E1, E2, C and their dimers). RVLP are secreted from the CHO24S cells into the medium and the time-point for collecting the medium with the highest yield of >100 kDa proteins (with 17 mg protein from 10 ml cell culture supernatant) was after 2 days of incubation. Different methods for RVLP isolation from the cell culture supernatants were assessed by SDS-PAGE and Western blotting (using sera positive or negative for Rubella virus (RV)-specific antibodies or an anti-E1 monoclonal antibody). A combination of membrane filtration with a rapid, novel gradient ultracentrifugation step (using Coomassie brilliant blue G crystals as adsorbens for RVLP that facilitated virus isolation) was the most suitable technique. 132 RV-positive human sera (RV IgG > 20 IU/ml by commercial ELISA) were tested by our "self made" immunoblot test stripes (using RVLP adsorbed to dye crystals as antigen) for the presence or absence of antibodies specific for RV structural proteins. 57.6% of these sera had antibodies against E1, E2 and C, 31% against E1 and C, and 1.5% against E1 only, whereas 3.8% had no RV specific antibodies and only 6.0% were equivocal which demonstrated that these "self made" test stripes can reliably differentiate RV antibody specificities.

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Detection of rubella virus-specific immunoglobulin M antibodies with a baculovirus-expressed E1 protein

Cited by 7 publications

References 40 publications

Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella

Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella

Baculovirus-Mediated Large-Scale Expression and Purification of a Polyhistidine-Tagged Rubella Virus Capsid Protein

Preparation of immunoblot test stripes from a Rubella virus-like particles dye crystal complex as antigen

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