Improving the diagnosis of 11β-hydroxylase deficiency using home-made MLPA probes: identification of a novel chimeric CYP11B2/CYP11B1 gene in a Sicilian patient
| Autor: | Laura Mazzanti, Antonio Balsamo, S Boccassini, Alessandra Gambineri, Soara Menabo, Lilia Baldazzi, Olga Prontera, Renato Pasquali |
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| Přispěvatelé: | Menabò, S, Boccassini, S, Gambineri, A, Balsamo, A, Pasquali, R, Prontera, O, Mazzanti, L, Baldazzi, L |
| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Adult Male Chimeric gene Prognosi Sequence analysis Endocrinology Diabetes and Metabolism 030209 endocrinology & metabolism 030105 genetics & heredity Homology (biology) 03 medical and health sciences Young Adult 0302 clinical medicine Endocrinology Medicine Cytochrome P-450 CYP11B2 Humans Multiplex Multiplex ligation-dependent probe amplification Gene Genetics Adrenal Hyperplasia Congenital business.industry Breakpoint Congenital adrenal hyperplasia Intron Sequence Analysis DNA Prognosis MLPA CYP11B1 11β-Hydroxylase deficiency Case-Control Studies Mutation Steroid 11-beta-Hydroxylase Female Case-Control Studie business Multiplex Polymerase Chain Reaction Human |
| Zdroj: | Journal of endocrinological investigation. 39(3) |
| ISSN: | 1720-8386 |
| Popis: | Purpose: 11β-Hydroxylase deficiency (11OHD) represents the second most common cause of congenital adrenal hyperplasia. It is caused by mutations in the CYP11B1 gene localized about 40 kb from the CYP11B2 gene with which it shares a homology of 95 %. The asymmetric recombination of these two genes is involved both in 11OHD and in glucocorticoid-remediable aldosteronism (GRA). Our objective was to set up an easy and rapid method to detect these hybrid genes and other kinds of deletions, to improve the molecular diagnosis of 11OHD. Methods: A set of 8 specific probes for both the CYP11B1 and the CYP11B2 genes to be used for multiplex ligation-dependent probe amplification (MLPA) analysis was designed to detect rearrangements of these genes. Results: The method developed was tested on 15 healthy controls and was proved to be specific and reliable; it led us to identify a novel chimeric CYP11B2/CYP11B1 gene in one patient that carried the known A306V mutation on the other allele. Specific amplification and sequencing of the hybrid gene confirmed the breakpoint localization in the second intron. Conclusions: The MLPA kit developed enables the detection of deletions, duplications or chimeric genes and represents an optimal supplement to DNA sequence analysis in patients with 11OHD. In addition, it can also be used to show the presence of the opposite chimaera associated with GRA. |
| Databáze: | OpenAIRE |
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