Analytical validation of a novel bioassay for thyroid-stimulating immunoglobulin.

Background: A novel and rapid cell-based bioassay, Turbo TSI, for measurement of thyroid-stimulating immunoglobulins (TSI) was recently reported. An assessment of the analytical performance of this TSI bioassay is described herein. Methods: Thawed cells from Turbo TSI kits were treated with different concentrations of a World Health Organization (WHO) international standard (IS) TSI-positive serum. TSI was measured as a function of luciferase activity measured as relative light units (RLU) and converted into international units per liter (IU/L). Analytical performance studies were performed on numerous samples, over multiple days, by two users at two sites. Results: The limit of blank, limit of detection and limit of quantitation were determined to be 0.007 IU/L, 0.014 IU/L, and 0.021 IU/L, respectively. Receiver operator characteristics (ROC) analysis determined the cut-off to be 0.0241 IU/L with an area under the curve of 0.984. The linear range was shown to be from 0.015 to 11.958 IU/L. The intra-laboratory precision was ≤15%CV. The overall reproducibility of the assay was ≤20%CV for five concentrations (0.06 to 5.16 IU/L). Interference and cross reactivity studies with a variety of substances showed that the assay was robust. The Turbo TSI bioassay demonstrated 95.2% (95% CI 83.3-98.1) positive percent agreement and 94.8% (95% CI 90.9-97.1) negative percent agreement with an FDA-cleared bioassay (Thyretain Conclusions: The Turbo TSI bioassay exhibits excellent analytical performance and a high level of reproducibility. The performance compared well with Thyretain