CYP21A2 Mutation Analysis in Korean Patients With Congenital Adrenal Hyperplasia Using Complementary Methods: Sequencing After Long-Range PCR and Restriction Fragment Length Polymorphism Analysis With Multiple Ligation-Dependent Probe Amplification Assay
Annals of Laboratory Medicine
; : 535-539, 2015.
Article
de En
| WPRIM
| ID: wpr-110960
Bibliothèque responsable:
WPRO
ABSTRACT
CYP21A2 mutation analysis of congenital adrenal hyperplasia (CAH) is challenging because of the genomic presence of a homologous CYP21A2 pseudogene and the significant incidence of pseudogene conversion and large deletions. The objective of this study was to accurately analyze the CYP21A2 genotype in Korean CAH patients using a combination of complementary methods. Long-range PCR and restriction fragment length polymorphism analyses were performed to confirm valid amplification of CYP21A2 and to detect large gene conversions and deletions before direct sequencing. Multiple ligation-dependent probe amplification (MLPA) analysis was conducted concurrently in 14 CAH-suspected patients and six family members of three patients. We identified 27 CYP21A2 mutant alleles in 14 CAH-suspected patients. The c.293-13A>G (or c.293-13C>G) was the most common mutation, and p.Ile173Asn was the second, identified in 25% and 17.9% of alleles, respectively. A novel frame-shift mutation of c.492delA (p.Glu 164Aspfs*24) was detected. Large deletions were detected by MLPA in 10.7% of the alleles. Mutation studies of the six familial members for three of the patients aided in the identification of haplotypes. In summary, we successfully identified CYP21A2 mutations using both long-range PCR and sequencing and dosage analyses. Our data correspond relatively well with the previously reported mutation spectrum analysis.
Mots clés
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Analyse spectrale
/
Haplotypes
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Polymorphisme de restriction
/
Pseudogènes
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Réaction de polymérisation en chaîne
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Incidence
/
Hyperplasie congénitale des surrénales
/
Allèles
/
Conversion des gènes
/
Génotype
Type d'étude:
Incidence_studies
/
Prognostic_studies
Limites du sujet:
Humans
Pays comme sujet:
Asia
langue:
En
Texte intégral:
Annals of Laboratory Medicine
Année:
2015
Type:
Article