Analytical validation of a novel bioassay for thyroid-stimulating immunoglobulin
BackgroundA 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.MethodsThawed cells from Turbo TSI kits were treated with different con...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Endocrinology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fendo.2024.1468768/full |
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| author | Paul D. Olivo Hannah Kim Lynn Miao Jeffery A. Houtz George J. Kahaly |
| author_facet | Paul D. Olivo Hannah Kim Lynn Miao Jeffery A. Houtz George J. Kahaly |
| author_sort | Paul D. Olivo |
| collection | DOAJ |
| description | BackgroundA 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.MethodsThawed 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.ResultsThe 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® TSI) using serum from 295 patients with autoimmune thyroid disease.ConclusionsThe Turbo TSI bioassay exhibits excellent analytical performance and a high level of reproducibility. The performance compared well with Thyretain® TSI, an FDA-cleared TSI bioassay. |
| format | Article |
| id | doaj-art-e52cd38c8141414593cc60f3b271ef39 |
| institution | DOAJ |
| issn | 1664-2392 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Endocrinology |
| spelling | doaj-art-e52cd38c8141414593cc60f3b271ef392025-08-20T02:43:38ZengFrontiers Media S.A.Frontiers in Endocrinology1664-23922025-01-011510.3389/fendo.2024.14687681468768Analytical validation of a novel bioassay for thyroid-stimulating immunoglobulinPaul D. Olivo0Hannah Kim1Lynn Miao2Jeffery A. Houtz3George J. Kahaly4Department of Microbiology and Microbial Pathogenesis, Washington University Medical School, St. Louis, MO, United StatesQuidelOrtho Corporation, San Diego, CA, United StatesQuidelOrtho Corporation, San Diego, CA, United StatesQuidelOrtho Corporation, San Diego, CA, United StatesMolecular Thyroid Research Laboratory, Department of Medicine I, Johannes Gutenberg University (JGU) Medical Center, Mainz, GermanyBackgroundA 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.MethodsThawed 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.ResultsThe 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® TSI) using serum from 295 patients with autoimmune thyroid disease.ConclusionsThe Turbo TSI bioassay exhibits excellent analytical performance and a high level of reproducibility. The performance compared well with Thyretain® TSI, an FDA-cleared TSI bioassay.https://www.frontiersin.org/articles/10.3389/fendo.2024.1468768/fullanalytical performancevalidationbioassaythyroid-stimulating receptor antibodythyroid-stimulating immunoglobulin |
| spellingShingle | Paul D. Olivo Hannah Kim Lynn Miao Jeffery A. Houtz George J. Kahaly Analytical validation of a novel bioassay for thyroid-stimulating immunoglobulin Frontiers in Endocrinology analytical performance validation bioassay thyroid-stimulating receptor antibody thyroid-stimulating immunoglobulin |
| title | Analytical validation of a novel bioassay for thyroid-stimulating immunoglobulin |
| title_full | Analytical validation of a novel bioassay for thyroid-stimulating immunoglobulin |
| title_fullStr | Analytical validation of a novel bioassay for thyroid-stimulating immunoglobulin |
| title_full_unstemmed | Analytical validation of a novel bioassay for thyroid-stimulating immunoglobulin |
| title_short | Analytical validation of a novel bioassay for thyroid-stimulating immunoglobulin |
| title_sort | analytical validation of a novel bioassay for thyroid stimulating immunoglobulin |
| topic | analytical performance validation bioassay thyroid-stimulating receptor antibody thyroid-stimulating immunoglobulin |
| url | https://www.frontiersin.org/articles/10.3389/fendo.2024.1468768/full |
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