Abstract |
Graves' disease is a type of autoimmune hyperthyroidism caused by thyroid-stimulating antibodies (TSAb).1 The combination of a porcine thyroid cell bioassay and cyclic adenosine monophosphate (cAMP) immunoassay (TSAb- enzyme immunoassay; EIA) is a clinically approved TSAb measurement method. Due to the requirement of multiple procedures and a long assay time of 6 h in the TSAb-EIA, a simplified and rapid assay is desired. Herein, we developed a rapid homogeneous TSAb bioassay (rapid-TSAb assay) using the human embryonic kidney cell line (HEK293), engineered to express the human thyroid-stimulating hormone receptor (TSHR), along with a cAMP-dependent luminescence biosensor. The measurement consists of three steps: thawing frozen cells, blood sample addition, and luminescence detection. The procedures can be conducted within 1 h. The World Health Organization International Standard TSAb (NIBSC 08/204) stimulated the cells co-expressing TSHR and cAMP biosensor. The intra- and inter-assay coefficients of variance were < 10%. Stimulation activity using wild-type TSHR and chimeric TSHR (Mc4) almost completely correlated with the tested Graves' disease and normal samples. In the rapid-TSAb assay, the evaluation of 39 samples, including TSHR antibody-positive sera, yielded a sensitivity of 100.0% and a specificity of 90.9%, compared to the TSAb-EIA control. The rapid-TSAb assay enables simple and rapid measurement of TSAb and is promising for improving the diagnosis of autoimmune thyroid diseases.
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Authors | Motoki Hoshina, Shiomi Ojima, Atsushi Kawasaki, Kosuke Doi, Satoshi Ohta, Asuka Inoue, Hiroshi Murayama |
Journal | Journal of immunological methods
(J Immunol Methods)
Vol. 528
Pg. 113655
(May 2024)
ISSN: 1872-7905 [Electronic] Netherlands |
PMID | 38447802
(Publication Type: Journal Article)
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Copyright | Copyright © 2023. Published by Elsevier B.V. |
Chemical References |
- Receptors, Thyrotropin
- Immunoglobulins, Thyroid-Stimulating
- Receptors, G-Protein-Coupled
- Thyrotropin
- Autoantibodies
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Topics |
- Animals
- Swine
- Humans
- Receptors, Thyrotropin
- HEK293 Cells
- Immunoglobulins, Thyroid-Stimulating
- Graves Disease
- Receptors, G-Protein-Coupled
- Thyrotropin
- Biological Assay
(methods)
- Autoantibodies
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