Abstract
Toxoplasmosis, while often an asymptomatic parasitic disease in healthy individuals, can cause severe complications in immunocompromised persons and during pregnancy. The most common method to diagnose Toxoplasma gondii infections is the serological determination of antibodies directed against parasite protein antigens. Here we report the use of a bead-based multiplex assay containing a synthetic phosphoglycan portion of the Toxoplasma gondii glycosylphosphatidylinositol (GPI1) for the detection of GPI1-specific antibodies in human sera. The glycan was conjugated to beads at the lipid site to retain its natural orientation and its immunogenic groups. We compared the response against GPI1 with that against the protein antigen SAG1, a common component of commercial serological assays, via the detection of parasite-specific human IgG and IgM antibodies, respectively. The GPI1-based test is in excellent agreement with the results for the commercial ELISA, as the ROC analysis of the GPI1 test shows 97% specificity and 98% sensitivity for the assay. GPI1 was a more reliable predictor for a parasite-specific IgM response compared to SAG1, indicating that a bead-based multiplex assay using GPI1 in combination with SAG1 may strengthen Toxoplasma gondii serology, in particular in seroepidemiological studies.
Highlights
T. gondii infections are primarily diagnosed by serological detection of IgM and IgG antibodies, and in some cases IgA, directed against parasitic protein antigens.[5]
GPI1 was a more reliable predictor for a parasite-specific IgM response compared to SAG1, indicating that a bead-based multiplex assay using GPI1 in combination with SAG1 may strengthen Toxoplasma gondii serology, in particular in seroepidemiological studies
Two main GPI glycoforms are present on the surface of T. gondii, a free GPI known as the low molecular weight antigen (GPI1),[13] and GPI2 that anchors proteins such as surface antigen SAG1 to the parasite membrane (Figure 1).[14,15]
Summary
T. gondii infections are primarily diagnosed by serological detection of IgM and IgG antibodies, and in some cases IgA, directed against parasitic protein antigens.[5]. Two main GPI glycoforms are present on the surface of T. gondii, a free GPI known as the low molecular weight antigen (GPI1),[13] and GPI2 that anchors proteins such as surface antigen SAG1 to the parasite membrane (Figure 1).[14,15] Synthetic GPI glycans fixed on array surfaces can be used to detect IgG and IgM anti-GPI antibodies in sera from infected individuals and to differentiate acute and latent toxoplasmosis infections.[16]. We report the use of a synthetic GPI-glycan conjugated to color-coded magnetic beads to detect anti-GPI1 antibodies using a BBMA This high-throughput method can simultaneously detect anti-SAG1 and anti-GPI1 antibodies with diagnostic value for toxoplasmosis and enable large-scale seroepidemiological studies.[17] our data show that comparing IgM responses directed against SAG1 and GPI1 could allow the discrimination between acute and postacute/latent infections
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