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The steroid response to human chorionic gonadotropin (hCG) stimulation in men with Klinefelter syndrome does not change using immunoassay or mass spectrometry

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Abstract

Purpose

Liquid-chromatography tandem mass-spectrometry (LC-MS/MS) was developed in parallel to Immunoassays (IAs) and today is proposed as the “gold standard” for steroid assays. Leydig cells of men with Klinefelter syndrome (KS) are able to respond to human chorionic gonadotropin (hCG) stimulation, even if testosterone (T) production was impaired. The aim was to evaluate how results obtained by IAs and LC-MS/MS can differently impact on the outcome of a clinical research on gonadal steroidogenesis after hCG stimulation.

Methods

A longitudinal, prospective, case-control clinical trial. (clinicaltrial.gov NCT02788136) was carried out, enrolling KS men and healthy age-matched controls, stimulated by hCG administration. Serum steroids were evaluated at baseline and for 5 days after intramuscular injection of 5000 IU hCG using both IAs and LC-MS/MS.

Results

13 KS patients (36 ± 9 years) not receiving T replacement therapy and 14 controls (32 ± 8 years) were enrolled. T, progesterone, cortisol, 17-hydroxy-progesterone (17OHP) and androstenedione, were significantly higher using IAs than LC-MS/MS. IAs and LC-MS/MS showed direct correlation for all five steroids, although the constant overestimation detected by IAs. Either methodology found the same 17OHP and T increasing profile after hCG stimulation, with equal areas under the curves (AUCs).

Conclusions

Although a linearity between IA and LC-MS/MS is demonstrated, LC-MS/MS is more sensitive and accurate, whereas IA shows a constant overestimation of sex steroid levels. This result suggests the need of reference intervals built on the specific assay. This fundamental difference between these two methodologies opens a deep reconsideration of what is needed to improve the accuracy of steroid hormone assays.

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Acknowledgements

DS is a PhD fellow of the Doctorate School in Clinical and Experimental Medicine of the University of Modena and Reggio Emilia, Italy. LC-MS/MS assays were performed thanks to Emilia-Romagna Region, Alessandro Liberati Young Researcher Grants, PRUA 1-2012-004.

Author information

Authors and Affiliations

  1. Department of Laboratory Medicine and Pathology Anatomy, Azienda USL of Modena, Modena, Italy

    L. Roli, S. Tagliavini, M. C. De Santis, E. Baraldi & T. Trenti

  2. Unit of Endocrinology, NOCSAE, Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, Via Giardini 1355, 41126, Modena, Italy

    D. Santi, S. Belli, V. Rochira, C. Carani & M. Simoni

  3. Department of Medicine, Endocrinology, Metabolism and Geriatrics, Azienda Ospedaliero-Universitaria of Modena, Modena, Italy

    D. Santi, A. R. Granata, V. Rochira & M. Simoni

  4. Laboratory of Clinical and Endocrinological Analysis, Arcispedale S. Maria Nuova, Reggio Emilia, Italy

    S. Cavalieri

  5. Endocrinology Unit and Centre for Applied Biomedical Research, Department of Medical and Surgical Sciences, Alma Mater University of Bologna, S. Orsola-Malpighi Hospital, 40138, Bologna, Italy

    F. Fanelli, M. Mezzullo, U. Pagotto & R. Pasquali

  6. Center for Genomic Research, University of Modena and Reggio Emilia, Modena, Italy

    M. Simoni

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  1. L. Roli
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Corresponding author

Correspondence to D. Santi.

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Conflict of interest

The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.

Ethical approval

The study protocol was approved by the Ethics Committee of Modena (File nr. 04/12).

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Written informed consent was obtained from all participants.

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Roli, L., Santi, D., Belli, S. et al. The steroid response to human chorionic gonadotropin (hCG) stimulation in men with Klinefelter syndrome does not change using immunoassay or mass spectrometry. J Endocrinol Invest 40, 841–850 (2017). https://doi.org/10.1007/s40618-017-0653-7

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