Congenital adrenal hyperplasia (CAH) is an autosomal recessive disease characterized by impaired adrenal steroidogenesis and most often caused by CYP21A2 gene mutations. For the first time, we reported complete spectrum and frequency of CYP21A2 gene mutations in 61 unrelated patients with classical and non-classical CAH from Serbia.
Direct DNA sequencing of whole CYP21A2 gene and polymerase chain reaction with sequence-specific primers for detection of CYP21A1P/CYP21A2 chimeras were combined.
We identified 18 different pathogenic alleles—two of them novel. Mutation detection rate was highest in patients with salt-wasting form of CAH (94.7 %). The most prevalent mutation was intron 2 splice site mutation, c.290-13A/C>G (18.5 %). Other mutation frequencies were: CYP21A1P/CYP21A2 chimeras (13 %), p.P30L (13 %), p.R356W (11.1 %), p.G110fs (7.4 %), p.Q318X (4.6 %), p.V281L (4.6 %), p.I172N (2.8 %), p.L307fs (2.8 %), p.P453S (1.9 %), etc. Mainly, frequencies were similar to those in Slavic populations and bordering countries. However, we found 6.5 % of alleles with multiple mutations, frequently including p.P453S. Effects of novel mutations, c.386T>C (p.Leu129Pro) and c.493T>C (p.Ser165Pro), were characterized in silico as deleterious. The effect of well-known mutations on Serbian patients’ phenotype was as expected.
The first comprehensive molecular genetic study of Serbian CAH patients revealed two novel CYP21A2 mutations. This study will enable genetic counseling in our population and contribute to better understanding of molecular landscape of CAH in Europe.
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Congenital adrenal hyperplasia
Cytochrome P450, family 21, subfamily A, polypeptide 2
Multiplex ligation-dependent probe amplification
Human gene mutation database
Non-classical form of CAH
Polymerase chain reaction
Sequence-specific PCR amplification
Simple virilizing form of CAH
Salt-wasting form of CAH
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Conflict of interest
The authors declare that they have no conflict of interest.
This study was funded by the Ministry of Education, Science and Technological Development, Republic of Serbia (Grant No. III41004).
This study involved human participants and has been approved by the Ethics Committee of the School of Medicine, University of Belgrade in Belgrade, Serbia and the Ethics Committee of the Mother and Child Health Care Institute of Serbia “Dr Vukan Cupic” in Belgrade, Serbia, and has therefore, been performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments.
Informed consent was obtained from all individual participants included in the study.
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Milacic, I., Barac, M., Milenkovic, T. et al. Molecular genetic study of congenital adrenal hyperplasia in Serbia: novel p.Leu129Pro and p.Ser165Pro CYP21A2 gene mutations. J Endocrinol Invest 38, 1199–1210 (2015). https://doi.org/10.1007/s40618-015-0366-8
- 21-Hydroxylase deficiency
- Alleles with multiple mutations
- CYP21A1P/CYP21A2 chimeras
- Genotype–phenotype correlation
- Mutation detection
- Mutations’ effect