| Texto completo | |
| Autor(es): |
Coeli-Lacchini, Fernanda Borchers
[1]
;
Turatti, Wendy
[1]
;
Lamparelli Elias, Paula Conde
[1]
;
Kagohara Elias, Lucila Leico
[2]
;
Martinelli, Jr., Carlos Eduardo
[3]
;
Moreira, Ayrton Custodio
[1]
;
Antonini, Sonir Roberto
[3]
;
de Castro, Margaret
[1]
Número total de Autores: 8
|
| Afiliação do(s) autor(es): | [1] Univ Sao Paulo, Sch Med Ribeirao Preto, Dept Internal Med, BR-14049 Ribeirao Preto, SP - Brazil
[2] Univ Sao Paulo, Sch Med Ribeirao Preto, Dept Physiol, BR-14049 Ribeirao Preto, SP - Brazil
[3] Univ Sao Paulo, Sch Med Ribeirao Preto, Dept Pediat, BR-14049 Ribeirao Preto, SP - Brazil
Número total de Afiliações: 3
|
| Tipo de documento: | Artigo Científico |
| Fonte: | Gene; v. 526, n. 2, p. 239-245, SEP 10 2013. |
| Citações Web of Science: | 5 |
| Resumo | |
Context: Molecular diagnosis of congenital adrenal hyperplasia (CAB) due to 21-hydroxylase deficiency (21OHD) has not been straightforward. Objective: To conduct a comprehensive genetic analysis by Multiplex Ligation dependent Probe Amplification (MLPA) and evaluate its reliability for the molecular CAI-21OHD diagnosis. Patients and methods: We studied 99 patients from 90 families with salt-wasting (SW; n = 32), simple-virilizing (SV; n = 29), and non-classical (NC; n = 29) CAH-21OHD. Molecular analysis was sequentially performed by detecting the most frequent point mutations by allele-specific oligonucleotide polymerase chain reaction (ASO-PCR), large rearrangements by MLPA, and rare mutations by direct sequencing. Parental segregation was evaluated. Results: ASO-PCR detected microconversions in 164 alleles (91.1%). MLPA identified CYP21A1P large conversions to CYP21A2 in 7 of the remaining 16 (43.7%), 30-kb deletions including the 3'-end of CYP21A1P, C4B, and the 5'-end of CYP21A2 in 3 of the 16 (18.7%), and a complete CYP21A2 deletion in one (63%). Five alleles (2.7%) required direct sequencing; three mutations located in the CYP21A2 gene and two derived from CYP21A1P were found. No parental segregation was observed in patients with the c.329\_336del and/or the CL6 cluster mutations. These cases were not diagnosed by ASO-PCR, but MLPA detected deletions in the promoter region of the CYP21A2 gene, explaining the genotype/phenotype dissociation. Conclusion: Using the proposed algorithm, all alleles were elucidated. False-positive results in MLPA occurred when mutations or polymorphisms were located close to the probe-binding regions. These difficulties were overcome by the association of MLPA with ASO-PCR and paternal segregation. Using these approaches, we can successfully use MLPA in a cost-effective laboratory routine for the molecular diagnosis of CAH-21OHD. (C) 2013 Elsevier B.V. All rights reserved. (AU) | |
| Processo FAPESP: | 07/58365-3 - Fisiopatologia e etiopatogenia molecular de doenças relacionadas aos eixos corticotrófico, somatotrófico e neurohipofisário |
| Beneficiário: | Margaret de Castro |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |