Androgens by immunoassay and mass spectrometry in children with 46,XY disorder of sex development

Abstract

Objective: Steroid measurement is a challenge in pediatric endocrinology. Currently, liquid chromatography with tandem mass spectrometry (LC-MS/MS) is considered a gold standard for this purpose. The aim of this study was to compare both LC-MS/MS and immunoassay (IA) for androgens before and after human recombinant chorionic gonadotropin (r-hCG) stimulus in children with 46,XY Disorders of Sex Development (DSD). Methods: Nineteen patients with 46,XY DSD were evaluated; all of them were prepubertal and non gonadectomized. Testosterone, dihydrotestosterone (DHT), dehydroepiandrosterone (DHEA) and androstenedione were measured by IA and LC-MS/MS before and 7 days after rhCG injection. The correlation between IA and LC-MS/MS was analysed by the intraclass correlation coefficient (ICC) and Spearman's rank correlation coefficient (SCC). For concordance analysis the Passing and Bablok (PB) regression and the Bland and Altman (BA) method were used. Results: Testosterone showed excellent correlation (ICC = 0.960 and SCC = 0.964); DHT showed insignificant and moderate correlations as indicated by ICC (0.222) and SCC (0.631), respectively; DHEA showed moderate correlation (ICC = 0.585 and SCC = 0.716); and androstenedione had poor and moderate correlations in ICC (0.363) and SCC (0.735), respectively. Using the PB method, all hormones showed a linear correlation, but proportional and systematic concordance errors were detected for androstenedione, systematic errors for testosterone and no errors for DHEA and DHT. By the BA method, there was a trend of IA to overestimate testosterone and androstenedione and underestimate DHEA and DHT when compared to LC-MS/MS. Conclusion: Traditional IA should be replaced by LC-MS/MS for the androgens measurement in prepubertal children whenever is possible.

Genetic Variants in SRD5A2 in a Spectrum of DSD Patients from Australian Clinics Highlight Importance of Genetic Testing alongside Typical First-Line Investigations

“…For one, age-appropriate reference ranges and recommended cut off values for T:DHT ratios are not well defined and can vary between different laboratories and countries [Bertelloni et al, 2018]. The methods used to measure androgens, typically performed by either immunoassay or mass spectrometry, may also affect the results, and although the latter is the gold standard, both methods are routinely used [Oliveira et al, 2020]. Additionally, hCG stimulation protocols can vary, which may influence the results (i.e., number of injections administered, timing of injections).…”

A prospective study on disorders of sex development in Duhok city, Kurdistan region, Iraq: Genetic, etiology and clinical presentation

“…A cooperative approach by a team consisting of an endocrinologist, a paediatrician, a surgeon, a radiologist, and a lab technician is necessary for the care of patients with DSD. Exterior genitalia virilization at birth is reliant on intrauterine exposure to androgens, which can come from testicles, adrenal glands, or occasionally exogenous sources (21). Mullerian structures generally indicate a genetic male sex or gonadal dysgenesis in a genetic male with insufficient anti-Mullerian hormone production (AMH).…”

Clinical utility of laboratory developed mass spectrometry assays for steroid hormone testing

“…A small percentage of testosterone is bound to cortisol-binding protein and alpha-1-acid glycoprotein, with the rest remaining free [ 1 ]. Clinically, total testosterone concentrations are measured in pediatric patients with ambiguous genitalia, precocious or delayed puberty; adult males with hypogonadism; and females with suspected polycystic ovarian syndrome, irregular menstrual periods, or androgen excess [ [2] , [3] , [4] ]. Further, in transgender individuals, testosterone may be measured to verify if gender-affirming therapy is successfully increasing or decreasing total testosterone concentrations [ [5] , [6] ].…”