Diagnosis of 21-hydroxylase deficiency by urinary metabolite ratios using gas chromatography-mass spectrometry analysis: Reference values for neonates and infants.
One major issue of newborn screening programs for 21-hydroxylase deficiency (21OHD) is the high rate of false-positive results, especially in preterm neonates. Urinary steroid metabolite analysis using gas chromatography-mass spectrometry (GC-MS) is suitable as a confirmatory diagnostic tool. The objective of this study was to analyze retrospectively diagnostic metabolite ratios in neonates and infants with and without 21OHD using GC-MS with emphasis on glucocorticoid metabolism, and to develop reference values for the steroid metabolite ratios for the diagnosis of 21OHD. We retrospectively analyzed urinary steroid hormone metabolites determined by GC-MS of 95 untreated neonates and infants with 21OHD (1-148 days), and 261 neonates and infants (100 preterms) without 21OHD (0-217 days). Metabolites of 17α-hydroxyprogesterone showed specificities below 98%, whereas the 21-deoxycortisol metabolite pregnanetriolone clearly separated 21OHD from non-21OHD subjects. The best diagnostic ratio for 21OHD was pregnanetriolone to 6α-hydroxy-tetrahydrocortisone. The lowest value of this ratio in the 21OHD group (0.47) was at least eight times higher than the highest values in the non-21OHD group (0.055). We have given appropriate reference values for steroid metabolite ratios in the largest 21OHD cohort so far described. Consideration of glucocorticoid metabolism, especially the use of typical neonatal 6α-hydroxylates metabolites, leads to improvement of diagnostic metabolite ratios.
Division of Pediatric Endocrinology and Diabetology, Laboratory for Translational Hormone Analysis in Pediatric Endocrinology, Steroid Research and Mass Spectromity Unit, Center of Child and Adolescent Medicine, Justus Liebig University, Giessen, Germany. Electronic address: Clemens.kamrath@paediat.med.uni-giessen.de.
Clemens
C
Kamrath
Division of Pediatric Endocrinology and Diabetology, Laboratory for Translational Hormone Analysis in Pediatric Endocrinology, Steroid Research and Mass Spectromity Unit, Center of Child and Adolescent Medicine, Justus Liebig University, Giessen, Germany.
Michaela F
MF
Hartmann
Division of Pediatric Endocrinology and Diabetology, Laboratory for Translational Hormone Analysis in Pediatric Endocrinology, Steroid Research and Mass Spectromity Unit, Center of Child and Adolescent Medicine, Justus Liebig University, Giessen, Germany.
Claudia
C
Boettcher
Division of Pediatric Endocrinology and Diabetology, Laboratory for Translational Hormone Analysis in Pediatric Endocrinology, Steroid Research and Mass Spectromity Unit, Center of Child and Adolescent Medicine, Justus Liebig University, Giessen, Germany.
Klaus-Peter
KP
Zimmer
Division of Pediatric Endocrinology and Diabetology, Laboratory for Translational Hormone Analysis in Pediatric Endocrinology, Steroid Research and Mass Spectromity Unit, Center of Child and Adolescent Medicine, Justus Liebig University, Giessen, Germany.
Stefan A
SA
Wudy
D000312:Adrenal Hyperplasia, Congenital / Q000175:diagnosis* / Q000378:metabolism / Q000652:urine*; D015331:Cohort Studies; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D055432:Metabolomics / Q000379:methods*; D011279:Pregnanetriol / Q000031:analogs & derivatives / Q000378:metabolism / Q000652:urine; D012016:Reference Values; D013256:Steroids / Q000378:metabolism / Q000652:urine; D013761:Tetrahydrocortisone / Q000031:analogs & derivatives / Q000378:metabolism / Q000652:urine
C017419:6 alpha-hydroxytetrahydrocortisone; D013256:Steroids; D011279:Pregnanetriol; D013761:Tetrahydrocortisone; C009030:pregnanetriolone
21-Hydroxylase deficiency; Congenital adrenal hyperplasia; Pregnanetriolone