The serum T3, T4, TSH, and free T4 levels were measured using the electro-chemiluminescence immunoassay (ECLIA) to assess the thyroid function. In the ECLIA, chemiluminescent ruthenium is used to electrically induce antigen-antibody reactions on the surface of steroptoavidin-coated paramagnetic micro-beads, and the luminescence emitted is measured to determine the concentration of the substance of interest. The Elecsys T3, Elecsys T4, Elecsys TSH, and Elecsys FT4 II kits from Roche Diagnostics Co., Ltd. were used. For the normal reference range, the normal range of each kit was used (range corresponding to the 2.5th–97.5th percentiles).
In addition, anti-thyroglobulin, anti-thyroid-stimulating hormone receptor antibody, and anti-thyroid peroxidase antibodies were measured in some patients with overt hypothyroidism.