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CC BY-NC-ND 4.0 · Nuklearmedizin 2021; 60(01): 38-46
DOI: 10.1055/a-1277-5972
Original Article

Third generation radioimmunoassay (RIA) for TSH receptor autoantibodies (TRAb) – one step less, similar results?

Drittgenerations – Radioimmunoassay (RIA) für TSH-Rezeptor-Autoantikörper (TRAK) – ein Schritt weniger, gleiche Ergebnisse?
Johannes J. Roggenbuck
1   Department of Nuclear Medicine, University Hospital Carl Gustav Carus, TU Dresden, Germany
,
Grit Zarske
2   Medipan GmbH, Berlin/Dahlewitz, Germany
,
Peter Schierack
3   Institute of Biotechnology, Faculty Environment and Natural Sciences, Brandenburg University of Technology, Senftenberg, Germany
,
Gerd Wunderlich
1   Department of Nuclear Medicine, University Hospital Carl Gustav Carus, TU Dresden, Germany
,
Karsten Conrad
4   Institute of Immunology, University Hospital Carl Gustav Carus, TU Dresden, Germany
,
Joerg Kotzerke
1   Department of Nuclear Medicine, University Hospital Carl Gustav Carus, TU Dresden, Germany
,
Dirk Roggenbuck   *
2   Medipan GmbH, Berlin/Dahlewitz, Germany
3   Institute of Biotechnology, Faculty Environment and Natural Sciences, Brandenburg University of Technology, Senftenberg, Germany
5   Faculty of Health Sciences, Joint Faculty of the Brandenburg University of Technology Cottbus – Senftenberg, the Brandenburg Medical School Theodor Fontane and the University of Potsdam, Germany
,
Klaus Zöphel   *
1   Department of Nuclear Medicine, University Hospital Carl Gustav Carus, TU Dresden, Germany
6   Department of Nuclear Medicine, Klinikum Chemnitz gGmbH, Chemnitz, Germany
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Abstract

Aim TSH-receptor (TSHR)-autoantibody (TRAb) is the serological hallmark of Graves’ disease (GD). Recently, 3rd-generation radioimmunoassays (RIA) employing monoclonal TRAb such as M22 or T7 instead of TSH for the inhibition of human TRAb binding with solid-phase TSHR (coated tubes) have been introduced into laboratory routine.

Methods As current assays typically employ a consecutive incubation of patient serum and labelled monoclonal TRAb, automation of TRAb RIA is a challenge. Thus, the assay procedure using human TSHR-coated tubes and the mouse monoclonal TRAb T7 was modified by combining both steps. The novel one-step method was compared with its corresponding consecutive 3rd-generation RIA by investigating 304 individuals encompassing 102 patients with active GD (GDa), 43 patients with GD after successful therapy (GDt), 31 with Hashimoto’s disease (HD), 28 with non-autoimmune thyroid diseases (NAITD) and 100 healthy subjects (HS).

Results With the new method, the incubation time was shortened by approximately one hour. Both 3rd-generation RIAs did not reveal a significantly different assay performance by comparing areas under the curve (AUC) with receiver operating characteristics curve analysis (AUC one-step: 0.94, AUC two-step: 0.96, p > 0.05, respectively). The two-step TRAb RIA demonstrated sensitivity and specificity values of 87.5 % and 96.2 %, respectively, whereas the one-step revealed 84.6 % and 96.2 %, respectively.

Conclusion One-step 3rd-generation RIA may be used for the reliable detection of TRAb. The shorter and easier assay design may improve its use and enable automation in routine nuclear medicine laboratories.

Zusammenfassung

Zielsetzung TSH-Rezeptor (TSHR)-Autoantikörper (TRAb) sind die pathogenetische Ursache des Morbus Basedow (GD). Die Bestimmung von TRAb in der Routinediagnostik ist mit Radioimmunoassays (RIA) der dritten Generation möglich. Diese verwenden monoklonale TRAb wie M22 oder T7 anstatt TSH, um die Bindung von TRAb an Festphasen-immobilisierte TSHR (coated tubes) zu inhibieren. Herkömmliche Assays lassen Patientenserum und markierte monoklonale TRAb nacheinander inkubieren. Dies erschwert eine Automatisierung des Assays.

Methoden Der Assay mit humanen TSHR-coated tubes und monoklonalem TRAb T7 wurde modifiziert, indem die beiden Schritte kombiniert wurden. Die neue Einschrittmethode wurde mit ihrem korrespondierenden RIA der dritten Generation verglichen. Eingeschlossen wurden 304 Individuen, davon 102 Patienten mit derzeit aktivem Morbus Basedow (GDa), 43 Patienten mit erfolgreich behandeltem Morbus Basedow (GDt), 31 Patienten mit Hashimoto-Thyreoiditis (HD), 28 Patienten mit nichtautoimmuner Schilddrüsenerkrankung (NAITD) und 100 gesunde Individuen (HS).

Ergebnisse Die Ablaufzeit des Assays konnte um etwa 1 Stunde verringert werden. Die beiden Assays unterschieden sich nicht signifikant voneinander bezüglich des Vergleichs der Lachen unter den Kurven (AUC) in der ROC-Analyse (AUC Einschrittmethode: 0,94; AUC Zweischrittmethode: 0,96; p > 0,05). Der herkömmliche TRAb-RIA zeigte jeweils eine Sensitivität und Spezifität von 87,5 % and 96,2 %, während die Werte des Einschrittassays bei 84,6 % und 96,2 % lagen.

Schlussfolgerung Die Einschrittmethode des RIA der dritten Generation kann für die verlässliche Bestimmung von TRAb verwendet werden. Das kürzere und einfachere Assaydesign kann die Anwendbarkeit der Methode verbessern und ihre Automatisierung in der nuklearmedizinischen Routinediagnostik ermöglichen.

Key words

thyroid - autoantibodies - TSH receptor autoantibody - Graves’ disease - Hashimoto’s thyroiditis

Schlüsselwörter

Schilddrüse - Autoantikörper - TSH-Rezeptor-Autoantikörper - Morbus Basedow - Hashimoto-Thyreoiditis

* These authors share senior authorship.




Publication History

Received: 25 February 2020

Accepted: 01 October 2020

Article published online:
03 February 2021

© 2021. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commecial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

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