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Immunoglobulin-like Domain of HsFcµR as a Capture Molecule for Detection of Crimean-Congo Hemorrhagic Fever Virus- and Zika Virus-Specific IgM Antibodies.

Rackow, Anne; Ehmen, Christa; von Possel, Ronald; Medialdea-Carrera, Raquel; Brown, David; Bispo de Filippis, Ana Maria; Carvalho de Sequeira, Patrícia; Ribeiro Nogueira, Rita Maria; Halili, Barie; Jakupi, Xhevat; Berisha, Lindita; Ahmeti, Salih; Sherifi, Kurtesh; Schmidt-Chanasit, Jonas; Schmitz, Herbert; Mika, Angela; Emmerich, Petra; Deschermeier, Christina.
Clin Chem; 65(3): 451-461, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30709812

BACKGROUND:

The cellular surface molecule HsTOSO/FAIM3/HsFcµR has been identified as an IgM-specific Fc receptor expressed on lymphocytes. Here, we show that its extracellular immunoglobulin-like domain (HsFcµR-Igl) specifically binds to IgM/antigen immune complexes (ICs) and exploit this property for the development of novel detection systems for IgM antibodies directed against Crimean-Congo hemorrhagic fever virus (CCHFV) and Zika virus (ZIKV).

METHODS:

His-tagged HsFcµR-Igl was expressed in Escherichia coli and purified by affinity chromatography, oxidative refolding, and size-exclusion chromatography. Specific binding of HsFcµR-Igl to IgM/antigen ICs was confirmed, and 2 prototypic ELISAs for the detection of anti-CCHFV and anti-ZIKV IgM antibodies were developed. Thereby, patient sera and virus-specific recombinant antigens directly labeled with horseradish peroxidase (HRP) were coincubated on HsFcµR-Igl-coated ELISA plates. Bound ICs were quantified by measuring turnover of a chromogenic HRP substrate.

RESULTS:

Assay validation was performed using paired serum samples from 15 Kosovar patients with a PCR-confirmed CCHFV infection and 28 Brazilian patients with a PCR-confirmed ZIKV infection, along with a panel of a priori CCHFV/ZIKV-IgM-negative serum samples. Both ELISAs were highly reproducible. Sensitivity and specificity were comparable with or even exceeded in-house gold standard testing and commercial kits. Furthermore, latex beads coated with HsFcµR-Igl aggregated upon coincubation with an IgM-positive serum and HRP-labeled antigen but not with either component alone, revealing a potential for use of HsFcµR-Igl as a capture molecule in aggregation-based rapid tests.

CONCLUSIONS:

Recombinant HsFcµR-Igl is a versatile capture molecule for IgM/antigen ICs of human and animal origin and can be applied for the development of both plate- and bead-based serological tests.
Selo DaSilva