The first study on the usefulness of recombinant tetravalent chimeric proteins containing fragments of SAG2, GRA1, ROP1 and AMA1 antigens in the detection of specific anti-Toxoplasma gondii antibodies in mouse and human sera
Autor: | Lucyna Holec-Gąsior, Weronika Grąźlewska, Dariusz Lautenbach, Katarzyna Dzitko, Bozena Dziadek, Bartłomiej Ferra, Justyna Gatkowska |
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Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
Serum Proteins Physiology Protozoan Proteins Antibodies Protozoan Biochemistry Serology Toxoplasma Gondii Mice Blood serum Immune Physiology Medicine and Health Sciences Enzyme-Linked Immunoassays Protozoans Multidisciplinary Immune System Proteins biology Eukaryota Recombinant Proteins Medicine Antibody Toxoplasma Toxoplasmosis Research Article Science Recombinant Fusion Proteins 030106 microbiology Immunology Antigens Protozoan Enzyme-Linked Immunosorbent Assay Research and Analysis Methods Sensitivity and Specificity Antibodies 03 medical and health sciences Antigen parasitic diseases medicine Parasitic Diseases Animals Humans Avidity Serologic Tests Antigens Immunoassays Protozoan Infections Organisms Toxoplasma gondii Biology and Life Sciences Proteins biology.organism_classification medicine.disease Fusion protein Virology Parasitic Protozoans 030104 developmental biology Immunoglobulin M Immunoglobulin G biology.protein Immunologic Techniques |
Zdroj: | PLoS ONE PLoS ONE, Vol 14, Iss 6, p e0217866 (2019) |
ISSN: | 1932-6203 |
Popis: | This study presents an evaluation of four tetravalent recombinant chimeric proteins containing fragments of the Toxoplasma gondii antigens, SAG2, GRA1, ROP1 and AMA1, as potential replacements of a the soluble, whole-cell tachyzoite lysate (TLA) used in serological assays. Recombinant chimeric proteins (SAG2-GRA1-ROP1-AMA1N, AMA1N-SAG2-GRA1-ROP1, AMA1C-SAG2-GRA1-ROP1, and AMA1-SAG2-GRA1-ROP1) obtained by genetic engineering were tested for their reactivity with specific IgM and IgG antibodies from sera of experimentally infected mice and humans with T. gondii infection using an enzyme-linked immunosorbent assay (ELISA). In total 192 serum samples from patients with acquired T. gondii infection and 137 sera from seronegative individuals were examined. The reactivity of chimeric antigens with antibodies generated during T. gondii invasion was measured and compared to the results obtained in assays based on whole-cell Toxoplasma antigen. Chimeric proteins proved effective in differentiation between T. gondii-infected and uninfected individuals (100% sensitivity and specificity in the IgG ELISAs) which shows their potential usefulness as a replacements for TLA in standardized commercial tests for the serodiagnosis of toxoplasmosis. In addition, the chimeric proteins were tested for use in avidity determination. Obtained results were comparable to those of the corresponding commercial assays, suggesting the utility of these proteins for avidity assessment. Furthermore, this study demonstrated that the AMA1-SAG2-GRA1-ROP1 chimeric protein has the potential to distinguish specific antibodies from serum samples of individuals with the early and chronic phase of T. gondii infection. |
Databáze: | OpenAIRE |
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