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.
10.3343/alm.2015.35.5.535
- Author:
Geehay HONG
1
;
Hyung Doo PARK
;
Rihwa CHOI
;
Dong Kyu JIN
;
Jae Hyeon KIM
;
Chang Seok KI
;
Soo Youn LEE
;
Junghan SONG
;
Jong Won KIM
Author Information
1. Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea. nayadoo@hanmail.net
- Publication Type:Brief Communication
- Keywords:
CYP21A2;
Pseudogene;
Restriction fragment length polymorphism;
Multiple ligation-dependent probe amplification;
Korea
- MeSH:
Adrenal Hyperplasia, Congenital*;
Alleles;
Gene Conversion;
Genotype;
Haplotypes;
Humans;
Incidence;
Korea;
Polymerase Chain Reaction*;
Polymorphism, Restriction Fragment Length*;
Pseudogenes;
Spectrum Analysis
- From:Annals of Laboratory Medicine
2015;35(5):535-539
- CountryRepublic of Korea
- Language:English
-
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.
