Salivary cortisol and cortisone in diagnosis of Cushing’s syndrome – a comparison of six different analytical methods

Nils Bäcklund; Göran Brattsand; Staffan Lundstedt; Elisabeth Aardal; Inga Bartuseviciene; Katarina Berinder; Charlotte Höybye; Pia Burman; Britt Edén Engström; Anders Isaksson; Anders Blomgren; Oskar Ragnarsson; Ulrika Rüetschi; Jeanette Wahlberg; Tommy Olsson; Per Dahlqvist

doi:10.1515/cclm-2023-0141

Published by De Gruyter April 4, 2023

Salivary cortisol and cortisone in diagnosis of Cushing’s syndrome – a comparison of six different analytical methods

Nils Bäcklund , Göran Brattsand , Staffan Lundstedt , Elisabeth Aardal , Inga Bartuseviciene , Katarina Berinder , Charlotte Höybye , Pia Burman , Britt Edén Engström , Anders Isaksson , Anders Blomgren , Oskar Ragnarsson , Ulrika Rüetschi , Jeanette Wahlberg , Tommy Olsson and Per Dahlqvist

From the journal Clinical Chemistry and Laboratory Medicine (CCLM)

https://doi.org/10.1515/cclm-2023-0141

Showing a limited preview of this publication:

Abstract

Objectives

Salivary cortisol and cortisone at late night and after dexamethasone suppression test (DST) are increasingly used for screening of Cushing’s syndrome (CS). We aimed to establish reference intervals for salivary cortisol and cortisone with three liquid chromatography-tandem mass spectrometry (LC-MS/MS) techniques and for salivary cortisol with three immunoassays (IAs), and evaluate their diagnostic accuracy for CS.

Methods

Salivary samples at 08:00 h, 23:00 h and 08:00 h after a 1-mg DST were collected from a reference population (n=155) and patients with CS (n=22). Sample aliquots were analyzed by three LC-MS/MS and three IA methods. After establishing reference intervals, the upper reference limit (URL) for each method was used to calculate sensitivity and specificity for CS. Diagnostic accuracy was evaluated by comparing ROC curves.

Results

URLs for salivary cortisol at 23:00 h were similar for the LC-MS/MS methods (3.4–3.9 nmol/L), but varied between IAs: Roche (5.8 nmol/L), Salimetrics (4.3 nmol/L), Cisbio (21.6 nmol/L). Corresponding URLs after DST were 0.7–1.0, and 2.4, 4.0 and 5.4 nmol/L, respectively. Salivary cortisone URLs were 13.5–16.6 nmol/L at 23:00 h and 3.0–3.5 nmol/L at 08:00 h after DST. All methods had ROC AUCs ≥0.96.

Conclusions

We present robust reference intervals for salivary cortisol and cortisone at 08:00 h, 23:00 h and 08:00 h after DST for several clinically used methods. The similarities between LC-MS/MS methods allows for direct comparison of absolute values. Diagnostic accuracy for CS was high for all salivary cortisol and cortisone LC-MS/MS methods and salivary cortisol IAs evaluated.

Keywords: Cushing’s syndrome; immunoassay; LC-MS/MS; method comparison; salivary cortisol; salivary cortisone

Corresponding author: Nils Bäcklund, Department of Public Health and Clinical Medicine, Umeå University, 901 87 Umeå, Sweden, Phone: +46706041730, Fax: +4690137633, E-mail: nils.backlund@umu.se

Funding source: Västerbotten Läns Landsting

Funding source: Private donation to Umeå University for research on pituitary disease

Acknowledgments

We would like to thank Katarina Iselid, Umeå University Hospital, for expert recruitment of reference individuals and sample management, and Anders Lundquist and Marie Eriksson, Umeå University, for statistical advice.

Research funding: This work was supported by the Västerbotten County Council and by a donation to Umeå University for research on pituitary disease.
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
Competing interests: Authors state no conflict of interest.
Informed consent: Informed consent was obtained from all individuals included in this study.
Ethical approval: Research involving human subjects complied with all relevant national regulations, institutional policies and is in accordance with the tenets of the Helsinki Declaration (as revised in 2013). This study was approved by the local Ethics Committee at Umeå University (Dnr 2015/08-31).

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/cclm-2023-0141).

Received: 2023-02-07

Accepted: 2023-03-21

Published Online: 2023-04-04

Published in Print: 2023-09-26