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Steroid biomarkers for identifying non-classic adrenal hyperplasia due to 21-hydroxylase deficiency in a population of PCOS with suspicious levels of 17OH-progesterone

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Abstract

Objective

We aimed at defining the most effective routine immunoassay- or liquid chromatography-tandem mass spectrometry (LC–MS/MS)-determined steroid biomarkers for identifying non-classic adrenal hyperplasia due to 21-hydroxylase deficiency (21-NCAH) in a PCOS-like population before genotyping.

Methods

Seventy PCOS-like patients in reproductive age with immunoassay-determined follicular 17OH-progesterone (17OHP) ≥ 2.00 ng/mL underwent CYP21A2 gene analysis and 1–24ACTH test. Serum steroids were measured by immunoassays at baseline and 60 min after ACTH stimulation; basal steroid profile was measured by LC–MS/MS.

Results

Genotyping revealed 23 21-NCAH, 15 single allele heterozygous CYP21A2 mutations (21-HTZ) and 32 PCOS patients displaying similar clinical and metabolic features. Immunoassays revealed higher baseline 17OHP and testosterone, and after ACTH stimulation, higher 17OHP (17OHP60) and lower cortisol, whereas LC–MS/MS revealed higher 17OHP (17OHPLC-MS/MS), progesterone and 21-deoxycortisol and lower corticosterone in 21-NCAH compared with both 21-HTZ and PCOS patients. Steroid thresholds best discriminating 21-NCAH from 21-HTZ and PCOS were estimated, and their diagnostic accuracy in identifying 21-NCAH from PCOS was established by ROC analysis. The highest accuracy was observed for 21-deoxycortisol ≥ 0.087 ng/mL, showing 100% sensitivity, while the combination of 17OHPLC-MS/MS ≥ 1.79 ng/mL and corticosterone ≤ 8.76 ng/mL, as well as the combination of ACTH-stimulated 17OHP ≥ 6.77 ng/mL and cortisol ≤ 240 ng/mL by immunoassay, showed 100% specificity.

Conclusions

LC–MS/MS measurement of basal follicular 21-deoxycortisol, 17OHP and corticosterone seems the most convenient method for diagnosing 21-NCAH in a population of PCOS with a positive first level screening, providing high accuracy and reducing the need for ACTH stimulation test.

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Acknowledgement

We thank Adrian Wallwork for English language revision.

Funding

This study was supported by the Emilia Romagna-Region, Alessandro Liberati Young Researcher Grant (Grant No. PRUA 1-2012-004).

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Author notes

  1. C. Oriolo and F. Fanelli shared first authorship.

Authors and Affiliations

  1. Endocrinology Unit and Centre for Applied Biomedical Research (CRBA), Department of Medical and Surgical Sciences, St. Orsola Hospital, University of Bologna, Via Massarenti 9, 40138, Bologna, Italy

    C. Oriolo, F. Fanelli, S. Castelli, M. Mezzullo, P. Altieri, F. Corzani, C. Pelusi, A. Repaci, G. Di Dalmazi, V. Vicennati, R. Pasquali, U. Pagotto & A. Gambineri

  2. Biostatistics Laboratory, Department of Medical and Surgical Sciences, St. Orsola Hospital, University of Bologna, Bologna, Italy

    A. Dormi, E. Nardi & G. Brillanti

  3. Medical Genetic Unit, Department of Medical and Surgical Sciences, St. Orsola Hospital, University of Bologna, Bologna, Italy

    L. Baldazzi & S. Menabò

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  1. C. Oriolo
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  2. F. Fanelli
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Correspondence to A. Gambineri.

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The study was approved by the independent ethical committee of Azienda Ospedaliero-Unversitaria of Bologna, Policlinico S. Orsola-Malpighi

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Oriolo, C., Fanelli, F., Castelli, S. et al. Steroid biomarkers for identifying non-classic adrenal hyperplasia due to 21-hydroxylase deficiency in a population of PCOS with suspicious levels of 17OH-progesterone. J Endocrinol Invest 43, 1499–1509 (2020). https://doi.org/10.1007/s40618-020-01235-3

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