Abstract
Purpose
Steroid 21-hydroxylase deficiency (21-OHD) is the common type of congenital adrenal hyperplasia (CAH) caused by defects in the CYP21A2 gene, as an autosomal recessive disease, genetic analysis has a prominent role in its diagnosis. Our objectives were to determine the prevalence of common mutations in a group of Egyptian patients with 21-OHD and their families using rapid methods, and also to detect the rate of deletion, duplication and conversions in CYP21A2 gene.
Methods
Rapid detection methods were used: allele-specific PCR for c.293-13A>G (g.659A>G), c.518T>A (p.I172N) variants and c.332_339del (8-bp deletion in exon 3), and real-time, quantitative PCR assay was used to detect deletion in the CYP21A2 gene. 29 Egyptian patients, 38 family members, and 20 healthy controls were all included in the study.
Results
The frequency of c.293-13A>G splice mutation was reported in 96.6 % cases, G allele had 2.5-folds higher risk to develop CAH than other alleles. The c.518T>A mutation was reported in 69 % cases, children carrying the mutant allele were 2.1 times more risk. The most frequent combined mutations detected were c.293-13A/C>G/c.518T>A in 58.6 % cases.
Conclusion
The genetic analysis of the splice site mutation c.293-13A>G and c.518T>A variant can be used as good biomarkers for early detection of cases and carriers in 21-OHD CAH Egyptian children, since the methods used have rapid turnaround time.
Similar content being viewed by others
References
Morel Y, Miller WL (1991) Clinical and molecular genetics of congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Adv Hum Genet 20:1–68
White PC, Tusie-Luna MT, New MI, Speiser PW (1994) Mutations in steroid 21-hydroxylase (CYP21). Hum Mut 3:373–378
New MI (1995) Steroid 21-hydroxylase deficiency (congenital adrenal hyperplasia). Am J Med 98(1A):S2–S8
Speiser PW (2004) Congenital adrenal hyperplasia. In: Pescovitz OH, Eugster EA (eds) Pediatric endocrinology: mechanism, manifestations and management. Lippincott Williams and Wilkins, Philadelphia, pp 600–613
Merke DP, Bornstein SR (2005) Congenital adrenal hyperplasia. Lancet 365:2125–2136
White PC, Speiser PW (2000) Congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Endocr Rev 21:245–291
Speiser PW (2001) Congenital adrenal hyperplasia owing to 21-hydroxylase deficiency. Endocrinol Metab Clin North Am 30:31–59
Speiser PW, Dupont B, Rubinstein P, Piazza A, Kastelan A, New MI (1985) High frequency of nonclassical steroid 21-hydroxylase deficiency. Am J Hum Genet 37:650–667
White PC, Grossberger D, Onufer BJ, Chaplin DD, New MI, Dupont B, Strominger JL (1985) Two genes encoding steroid 21-hydroxylase are located near the genes encoding he fourth component of complement in man. Proc Natl Acad Sci USA 82:1089–1093
Higashi Y, Yoshioka H, Yamane M, Gotoh O, Fujii-Kuriyama Y (1986) Complete nucleotide sequence of two steroid 21-hydroxylase genes tandemly arranged in human chromosome: a pseudogene and a genuine gene. Proc Natl Acad Sci USA 83:2841–2845
Nimkarn SI, New MI (2007) Prenatal diagnosis and treatment of congenital adrenal hyperplasia owing to 21-hydroxylase deficiency. Nat Clin Pract Endocrinol Metab 3:405–413
White PC, New MI, Dupont B (1986) Structure of human steroid 21-hydroxylase genes. Proc Natl Acad Sci USA 83:5111–5115
Forest MG (2004) Recent advances in diagnosis and management of congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Hum Reprod Update 10:469–485
Haider S, Islam B, D’Atri V, Sgobba M, Poojari C, Sun L, Yuen T, Zaidi M, New MI (2013) Structure-phenotype correlations of human CYP21A2 mutations in congenital adrenal hyperplasia. Proc Natl Acad Sci USA 12(110):2605–2610
Helmberg A, Tusie-Luna MT, Tabarelli M, Kofler R, White PC (1992) R339H and P453S: CYP21 mutations associated with nonclassic steroid 21-hydroxylase deficiency that are not apparent gene conversions. Mol Endocrinol 6:1318–1322
Temtamy S, Aglan M (2012) Consanguinity and genetic disorders in Egypt. Middle East J Med Genet 1:12–17
Speiser PW, Azziz R, Baskin LS, Ghizzoni L, Hensle TW, Merke DP, Meyer-Bahlburg HF, Miller WL, Montori VM, Oberfield SE, Ritzen M, White PC (2010) Endocrine society: congenital adrenal hyperplasia due to steroid 21 hydroxylase deficiency: an endocrine society clinical practice guideline. J Clin Endocrinol Metab 95:4133–4160
Newton CR, Graham A, Heptinstall LE, Powell SJ, Summers C, Kalsheker N, Smith JC, Markham AF (1989) Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). Nucleic Acids Res 17:2503–2516
Krone N, Braun A, Weinert S, Peter M, Roscher A, Partsch C (2002) Multiplex minisequencing of the 21-hydroxylase gene as a rapid strategy to confirm congenital adrenal hyperplasia. Clin Chem 48:818–825
Weisenpacherová R, Pribilincová Z, Behulová R, Mezenská R, Lukáčová M (2004) Mutational analysis of CYP21 gene in Slovak patients with 21-hydroxylase deficiency and comparison with other European populations. Biol Bratisl 59:795–802
Olney R, Mougey E, Wang J, Shulman D, Sylvester J (2002) Using real-time, quantitative PCR for rapid genotyping of the steroid 21-hydroxylase gene in a north Florida population. J Clin Endocrinol Metab 87:735–741
Lee C, Tung Y, Hsiao P, Lee J, Tsai W (2010) Clinical characteristics of Taiwanese children with congenital adrenal hyperplasia caused by 21-hydroxylase deficiency in the pre-screening era. J Formos Med Assoc 109:148–155
Speiser PW, White PC (2003) Congenital adrenal hyperplasia. N Engl J Med 21(349):776–788
Wedell A (1998) Molecular genetics of congenital adrenal hyperplasia (21-hydroxylase deficiency): implications for diagnosis, prognosis and treatment. Acta Paediatr 87:159–164
New MI, Abraham M, Gonzalez B, Dumic M, Razzaghy-Azar M, Chitayat D, Sun L, Zaidi M, Wilson RC, Yuen T (2013) Genotype-phenotype correlation in 1,507 families with congenital adrenal hyperplasia owing to 21-hydroxylase deficiency. Proc Natl Acad Sci USA 12(110):2611–2616
Menabò S, Balsamo A, Baldazzi L, Barbaro M, Nicoletti A, Conti V, Pirazzoli P, Wedell A, Cicognani A (2012) A sequence variation in 3′UTR of CYP21A2 gene correlates with a mild form of congenital adrenal hyperplasia. J Endocrinol Invest 35:298–305
Khan AH, Aban M, Raza J, Ul Haq N, Jabbar A, Moatter T (2011) Ethnic disparity in 21-hydroxylase gene mutations identified in Pakistani congenital adrenal hyperplasia patients. BMC Endocr Disord 11:5
Balraj P, Lim PG, Sidek H, Wu LL, Khoo AS (2013) Mutational characterization of congenital adrenal hyperplasia due to 21-hydroxylase deficiency in Malaysia. J Endocrinol Invest. 36:366–374
Torres N, Mello M, Germano C, Elias L, Moreira M, Castro M (2003) Phenotype and genotype correlation of the microconversion from the CYP21A1P to the CYP21A2 gene in congenital adrenal hyperplasia. Braz J Med Biol Res 36:1311–1318
Bobba A, Marra E, Giannattasio S, Iolascon A, Monno F (1999) 21-hydroxylase deficiency in Italy: a distinct distribution pattern of CYP21 mutations in a sample from southern Italy. J Med Genet 36:648–650
White P, Speiser P (2000) Congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Endocr Rev 21:245–291
Dolzan V, Sólyom J, Fekete G, Kovács J, Rakosnikova V, Votava F, Lebl J, Pribilincova Z, Baumgartner-Parzer SM, Riedl S, Waldhauser F, Frisch H, Stopar-Obreza M, Krzisnik C, Battelino T (2005) Mutational spectrum of steroid 21-hydroxylase and the genotype-phenotype association in Middle European patients with congenital adrenal hyperplasia. Eur J Endocrinol 153:99–106
Conflict of interest
All authors declare that there is no commercial affiliation or consultancy of any of the authors that could be construed as a conflict of interest with respect to the submitted data.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sharaf, S., Hafez, M., ElAbd, D. et al. High frequency of splice site mutation in 21-hydroxylase deficiency children. J Endocrinol Invest 38, 505–511 (2015). https://doi.org/10.1007/s40618-014-0207-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s40618-014-0207-1