Abstract
A sensitive nonisotopic immunoassay for the determination of 17-hydroxyprogesterone (17-OHP) levels in saliva was developed. The new time-resolved fluorometric immunoassay employs a specific polyclonal anti-17-OHP antiserum immobilized onto microtiter plates, a 17-OHP-biotin conjugate as a tracer, and streptavidin-europium a as secondary probe. The lower detection limit of the assay is 23.6 pmol/L (mean -3 s of a 22-fold zero determination) corresponding to 0.39 pg/well. The coefficients of intraassay variation are 8.8, 5.3, and 8.3% at the respective concentrations of 90.9, 454.5, and 1363.5 pmol/L. The coefficients of interassay variation are 8.8, 5.3, and 8.3% at the respective concentrations. Saliva was collected in commercially available devices. Reference ranges were established using 394 saliva samples from 132 healthy children, adolescents, and adults. Morning, midday, and evening levels of 17-OHP levels in saliva varied significantly in all age groups with morning levels being higher than midday and evening levels. Saliva samples (n = 57) were also obtained from 18 children with congenital adrenal hyperplasia (CAH). Salivary 17-OHP levels in the limited number of CAH patients studied ranged from 121 to 106 050 pmol/L. In conclusion 1) a new, sensitive nonisotopic immunoassay for measurement of 17-OHP in saliva has been developed; 2) reference ranges for healthy children, adolescents, and adults have been established; 3) there is a circadian pattern of 17-OHP levels in saliva at all ages; and 4) measurement of 17-OHP in saliva should be further evaluated over a longer period of time as a potentially reliable and powerful technique to monitor metabolic control in patients with CAH. As 17-OHP levels in saliva are stable for > 10 wk at 4°C, the technique is ideally suited for outpatient sampling.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- CAH:
-
congenital adrenal hyperplasia
- TR-FIA:
-
time-resolved fluorometric immunoassay
- 17-OHP:
-
17-hydroxyprogesterone
- 17-OHP-3CMO:
-
17-hydroxyprogesterone-3-O-carboxymethyloxime
References
New MI 1995 Steroid 21-hydroxylase deficiency (congenital adrenal hyperplasia). Am J Med 98: 2S–8S
White PC, New MI 1992 Genetic basis of endocrine disease: congenital adrenal hyperplasia due to 21-hydroxylase deficiency. J Clin Endocrinol Metab 74: 6–11
Killeen AA, Hanson NQ, Eklund R, Vairl CJ, Eckfeldt JH 1992 Prevalence of nonclassical congenital adrenal hyperplasia among women self-referred for electrolytic treatment of hirsutism. Am J Med Genet 42: 197–200
James VH, Few JD 1985 Adrenocorticosteroids: chemistry, synthesis and disturbances in disease. Clin Endocrinol Metab 14: 867–892
Lim YJ, Batch JA, Warne GL 1995 Adrenal 21-hydroxylase deficiency in childhood: 25 years' experience. J Paediatr Child Health 31: 222–227
Rittner HL, Lee PDK, Blum WF, Doerr HG, Steiss J, Kreuder J, Rascher W, Kiess W 1997 Developmental patterns of serum 3α-androstandiol glucuronide. J Endocrinol Invest 20: 245–250
Otten BJ, Wellen JJ, Rijen JC, Stoelinga GB, Benraad TJ 1983 salivary and plasma androstenedione and 17-hydroxyprogesterone levels in congenital adrenal hyperplasia. J Clin Endocrinol Metab 57: 1150–1154
Von Schnakenburg K, Bidlingmaier F, Knorr D 1980 17-hydroxyprogesterone, androstenedione, and testosterone in normal children and in prepubertal patients with congenital adrenal hyperplasia. Eur J Pediatr 133: 259–267
Zerah M, Pang S, New MI 1987 Morning salivary 17-hydroxyprogesterone is a useful screening test for nonclassical 21-hydroxylase deficiency. J Clin Endocrinol Metab 65: 227–232
Zerah M, Ueshiba H, Wood E, Speiser PW, Crawford C, McDonald T, Pareira J, Gruen D, New MI 1990 Prevalence of nonclassical steroid 21-hydroxylase deficiency based on a morning salivary 17-OHP screening test: a small sample study. J Clin Endocrinol Metab 70: 1662–1667
Hampl R, Foretova L, Sulcova J, Starka L 1990 Daily profiles of salivary cortisol in hydrocortisone treated children with congenital adrenal hyperplasia. Eur J Pediatr 149: 232–234
Walker RF, Hughes IA, Riad-Fahmy D 1979 Salivary 17α-OHP in congenital adrenal hyperplasia. Clin Endocrinol 11: 631–637
Price DA, Astin MP, Chard CR, Addison GM 1979 Assay of hydroxyprogesterone in saliva. Lancet 2: 368
O'Rorke A, Kane MM, Gosling JP, Tallon DF, Fottrell PF 1994 Development and validation of a monoclonal antibody enzyme immunoassay for measuring progesterone in saliva. Clin Chem 40: 454–458
Shen SX, Young MC, Hinohosa-Sandoval M, Hughes IA 1989 17-OH-Progesterone response to acute dexamethasone administration in congenital adrenal hyperplasia. Horm Res 32: 136–141
Young MC, Robinson JA, Read GF, Riad-Fahmy D, Hughes IA 1988 17-OH-Progesterone rhythms in congenital adrenal hyperplasia. Arch Dis Child 63: 617–623
Nahoul K, Scholler R 1987 Comparison of saliva and plasma 17-hydroxyprogesterone time-course response to hCG administration in normal men. J Steroid Biochem 26: 251–257
Hoepffner W, Hubl W 1986 Studies on the diurnal variations of 17-hydroxyprogesterone in saliva by enzyme immunoassay in patients with congenital adrenal hyperplasia. Exp Clin Endocrinol 87: 189–194
Hughes IA, Days J, Robinson J, Walker RF, Riad-Fahmy D 1985 Monitoring treatment in congenital adrenal hyperplasia. Use of serial measurements of 17-OH-progesterone in plasma, capillary blood, and saliva. Ann NY Acad Sci 458: 193–202
Hughes IA, Read GF 1984 Control in congenital adrenal hyperplasia monitored by frequent saliva 17-OH-progesterone measurements. Horm Res 19: 77–85
Dyas J, Read GF, Guha-Maulik T, Hughes IA, Riad-Fahmy D 1984 A rapid assay for 17α-OH-progesterone in plasma, saliva and amniotic fluid using a magnetisable solid-phase antiserum. Ann Clin Biochem 21: 417–424
Price DA, Astin MP, Chard CR, Addison GM 1979 Assay of hydroxyprogesterone in saliva. Lancet 2: 368–369
Walker RF, Hughes IA, Riad-Fahmy D 1979 Salivary 17-α-OH-progesterone in congenital adrenal hyperplasia. Clin Endocrinol 11: 631–637
Walker RF, Read GF, Hughes IA, Riad-Fahmy D 1979 Salivary 17-α-OH-progesterone in congenital adrenal hyperplasia. Clin Chem 25: 542–545
Walker RF, Fahmy DR 1978 Radioimmunoassay of 17α-hydroxyprogesterone in whole saliva and parotid fluid of children with congenital hyperplasia. J Endocrinol 79: 64P–65P
Hammond GL, Langley MS 1986 Identification and measurement of SHBG and CBG in human saliva. Acta Endocrinol 112: 603–608
Price DA 1984 Salivary hormone levels in infants and children. Front Oral Physiol 5: 51–68
Vining RF, McGinley RA 1984 Flux of steroids between blood and saliva. Front Oral Physiol 5: 21–22
Luisi M, Franchi F 1984 Salivary steroid measurement. An alternative approach to plasma assays in assessing endocrine function. Front Oral Physiol 5: 124–154
Riad-Fahmy D, Read GF, Walker RF, Griffiths K 1982 Steroids in saliva for assessing endocrine function. Endocr Rev 3: 367–395
Kiess W, Meidert A, Dressendorfer RA, Schriever K, Kessler U, Konig A, Schwarz HP, Strasburger CJ 1995 Salivary cortisol levels throughout childhood and adolescence: relation with age, pubertal stage, and weight. Pediatr Res 37: 502–506
Kirschbaum C 1991 Cortisolmessung im Speichel-Eine Methode der Biologischen Psychologie. Verlag Hans Huber, Toronto, pp 32–70.
Ueshiba H, Zerah M, New MI 1994 Enzyme-linked immunosorbent assay (ELISA) method for screening of non-classical steroid 21-hydroxylase deficiency. Horm Metab Res 26: 43–45
Dressendorfer R, Kirschbaum C, Rohde W, Stahl F, Strasburger CJ 1992 Synthesis of a cortisol-biotin conjugate and evaluation as a tracer in an immunoassay for salivary cortisol measurement. J Steroid Biochem Mol Biol 43: 683–692
Meulenberg EP, Hofman JA 1990 The effect of pretreatment of saliva on steroid hormone concentrations. J Clin Chem Clin Biochem 28: 923–928
Laue L, Merke DP, Jones JV, Barnes KM, Hill S, Cutler GB 1996 A preliminary study of flutamide, testolactone, and reduced hydrocortisone dose in the treatment of congenital adrenal hyperplasia. J Clin Endocrinol Metab 81: 3535–3539
Van Wyk JJ, Gunther DF, Ritzen EM, Wedell A, Cutler GB, Migeon CJ, New MI 1996 The use of adrenalectomy as a treatment for congenital adrenal hyperplasia. J Clin Endocrinol Metab 81: 3180–3190
Acknowledgements
The authors thank Christian Smit for help in collecting samples and Professor Dr. Wolfgang Rascher for his support. We express our sincere gratitude to the children and their families who participated in the study. We gratefully acknowledge the generous gift of saliva collecting devices (Salivette®) by Sarstedt, Numbrecht, Germany.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Dressendorfer, R., Strasburger, C., Bidlingmaier, F. et al. Development of a Highly Sensitive Nonisotopic Immunoassay for the Determination of Salivary 17-Hydroxyprogesterone: Reference Ranges throughout Childhood and Adolescence. Pediatr Res 44, 650–655 (1998). https://doi.org/10.1203/00006450-199811000-00006
Received:
Accepted:
Issue date:
DOI: https://doi.org/10.1203/00006450-199811000-00006
This article is cited by
-
17-Hydroxyprogesteron im Speichel
Monatsschrift Kinderheilkunde (2012)
