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Molecular CYP21A2 diagnosis in 480 Brazilian patients with congenital adrenal hyperplasia before newborn screening introduction.
Eur J Endocrinol. 2016 Aug; 175(2):107-16.EJ

Abstract

BACKGROUND

Most congenital adrenal hyperplasia (CAH) patients carry CYP21A2 mutations derived from conversion events involving the pseudogene, and the remaining carry new mutations.

OBJECTIVE

To review causal mutations and genotype-phenotype correlation in 480 Brazilian patients.

METHODS

DNA was extracted from 158 salt-wasters (SWs), 116 simple virilizing (SV), and 206 nonclassical (NC) patients. Fourteen point mutations were screened by allele-specific PCR, large rearrangements by Southern blotting/MLPA, and sequencing was performed in those with incomplete genotype. The gene founder effect was analyzed by microsatellite studies. Patients were divided into six genotypes (Null; A: <2%; B: 3-7%; C: >20% of residual enzymatic activity (EA); D: unknown EA; E: incomplete genotype).

RESULTS

Targeted methodologies defined genotype in 87.6% of classical and in 80% of NC patients and the addition of sequencing in 100 and 83.5%, respectively. The most frequent mutations were p.V281L (26.6% of alleles), IVS2-13A/C>G (21.1%), and p.I172N (7.5%); seven rare mutations and one novel mutation (p.E351V) were identified. Gene founder effect was observed in all but one (p.W19X) mutation. Null, A, B, and C genotypes correlated with SW (88%), SW (70%), SV (98%), and NC forms (100%), respectively. In group D, the p.E351V mutation correlated with classical form and group E comprised exclusively NC-patients. ACTH-stimulated 17OHP level of 44.3ng/mL was the best cutoff to identify NC-patients carrying severe mutations.

CONCLUSIONS

We identified a good genotype-phenotype correlation in CAH, providing useful data regarding prediction of disease's severity; moreover, we suggest that ACTH-stimulated 17OHP levels could predict carrier status for severe mutations. Sequencing is essential to optimize molecular diagnosis in Brazilian CAH patients.

Authors+Show Affiliations

Laboratório de Hormônios e Genética Molecular- LIM/42Unidade de Adrenal, Disc. de Endocrinologia e Metabologia dacarv89@gmail.com.Laboratório de Hormônios e Genética Molecular- LIM/42Unidade de Adrenal, Disc. de Endocrinologia e Metabologia.Laboratório de Hormônios e Genética Molecular- LIM/42Unidade de Adrenal, Disc. de Endocrinologia e Metabologia.Laboratório de Hormônios e Genética Molecular- LIM/42Unidade de Adrenal, Disc. de Endocrinologia e Metabologia.Laboratório de Hormônios e Genética Molecular- LIM/42Unidade de Adrenal, Disc. de Endocrinologia e Metabologia.Laboratório de Hormônios e Genética Molecular- LIM/42Unidade de Adrenal, Disc. de Endocrinologia e Metabologia.Laboratório de Hormônios e Genética Molecular- LIM/42Unidade de Adrenal, Disc. de Endocrinologia e Metabologia.Unidade de Endocrinologia PediátricaInstituto da Criança, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil.Unidade de Endocrinologia PediátricaInstituto da Criança, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil.Unidade de Endocrinologia PediátricaInstituto da Criança, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil.Laboratório de Hormônios e Genética Molecular- LIM/42Unidade de Adrenal, Disc. de Endocrinologia e Metabologia.Laboratório de Hormônios e Genética Molecular- LIM/42Unidade de Adrenal, Disc. de Endocrinologia e Metabologia.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27185867

Citation

de Carvalho, Daniel F., et al. "Molecular CYP21A2 Diagnosis in 480 Brazilian Patients With Congenital Adrenal Hyperplasia Before Newborn Screening Introduction." European Journal of Endocrinology, vol. 175, no. 2, 2016, pp. 107-16.
de Carvalho DF, Miranda MC, Gomes LG, et al. Molecular CYP21A2 diagnosis in 480 Brazilian patients with congenital adrenal hyperplasia before newborn screening introduction. Eur J Endocrinol. 2016;175(2):107-16.
de Carvalho, D. F., Miranda, M. C., Gomes, L. G., Madureira, G., Marcondes, J. A., Billerbeck, A. E., Rodrigues, A. S., Presti, P. F., Kuperman, H., Damiani, D., Mendonca, B. B., & Bachega, T. A. (2016). Molecular CYP21A2 diagnosis in 480 Brazilian patients with congenital adrenal hyperplasia before newborn screening introduction. European Journal of Endocrinology, 175(2), 107-16. https://doi.org/10.1530/EJE-16-0171
de Carvalho DF, et al. Molecular CYP21A2 Diagnosis in 480 Brazilian Patients With Congenital Adrenal Hyperplasia Before Newborn Screening Introduction. Eur J Endocrinol. 2016;175(2):107-16. PubMed PMID: 27185867.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular CYP21A2 diagnosis in 480 Brazilian patients with congenital adrenal hyperplasia before newborn screening introduction. AU - de Carvalho,Daniel F, AU - Miranda,Mirela C, AU - Gomes,Larissa G, AU - Madureira,Guiomar, AU - Marcondes,José A M, AU - Billerbeck,Ana Elisa C, AU - Rodrigues,Andresa S, AU - Presti,Paula F, AU - Kuperman,Hilton, AU - Damiani,Durval, AU - Mendonca,Berenice B, AU - Bachega,Tania A S S, Y1 - 2016/05/16/ PY - 2016/02/23/received PY - 2016/05/16/accepted PY - 2016/5/18/entrez PY - 2016/5/18/pubmed PY - 2017/2/7/medline SP - 107 EP - 16 JF - European journal of endocrinology JO - Eur J Endocrinol VL - 175 IS - 2 N2 - BACKGROUND: Most congenital adrenal hyperplasia (CAH) patients carry CYP21A2 mutations derived from conversion events involving the pseudogene, and the remaining carry new mutations. OBJECTIVE: To review causal mutations and genotype-phenotype correlation in 480 Brazilian patients. METHODS: DNA was extracted from 158 salt-wasters (SWs), 116 simple virilizing (SV), and 206 nonclassical (NC) patients. Fourteen point mutations were screened by allele-specific PCR, large rearrangements by Southern blotting/MLPA, and sequencing was performed in those with incomplete genotype. The gene founder effect was analyzed by microsatellite studies. Patients were divided into six genotypes (Null; A: <2%; B: 3-7%; C: >20% of residual enzymatic activity (EA); D: unknown EA; E: incomplete genotype). RESULTS: Targeted methodologies defined genotype in 87.6% of classical and in 80% of NC patients and the addition of sequencing in 100 and 83.5%, respectively. The most frequent mutations were p.V281L (26.6% of alleles), IVS2-13A/C>G (21.1%), and p.I172N (7.5%); seven rare mutations and one novel mutation (p.E351V) were identified. Gene founder effect was observed in all but one (p.W19X) mutation. Null, A, B, and C genotypes correlated with SW (88%), SW (70%), SV (98%), and NC forms (100%), respectively. In group D, the p.E351V mutation correlated with classical form and group E comprised exclusively NC-patients. ACTH-stimulated 17OHP level of 44.3ng/mL was the best cutoff to identify NC-patients carrying severe mutations. CONCLUSIONS: We identified a good genotype-phenotype correlation in CAH, providing useful data regarding prediction of disease's severity; moreover, we suggest that ACTH-stimulated 17OHP levels could predict carrier status for severe mutations. Sequencing is essential to optimize molecular diagnosis in Brazilian CAH patients. SN - 1479-683X UR - https://www.unboundmedicine.com/medline/citation/27185867/Molecular_CYP21A2_diagnosis_in_480_Brazilian_patients_with_congenital_adrenal_hyperplasia_before_newborn_screening_introduction_ L2 - https://eje.bioscientifica.com/doi/10.1530/EJE-16-0171 DB - PRIME DP - Unbound Medicine ER -