![]() ![]() Ammini, Molecular genetic analysis of CYP21A2 gene in patients with congenital adrenal hyperplasia. Waxman, The P450 superfamily: update on new sequences, gene mapping, and recommended nomenclature. Yuen, Genotype-phenotype correlation in 1,507 families with congenital adrenal hyperplasia owing to 21-hydroxylase deficiency. National Family Health Survey (NFHS-4), 2015-16. International Institute for Population Sciences (IIPS) and ICF. Atwal, Current and future perspective of newborn screening: an Indian scenario. Fujieda, Newborn screening for congenital adrenal hyperplasia in Sapporo City: sixteen years’ experience. Clark, Newborn screening, prenatal diagnosis and prenatal treatment of congenital adrenal hyperplasia due to 21-hydroxylase deficiency. We describe CYP21A2 mutation spectrum and novel variants in a large cohort of Indian patients with CAH.ĭ. ![]() We have developed and validated a comprehensive NGS-based assay for detection of variants in CYP21A2 gene in patients with 21-OH CAH. Thirty samples tested orthogonally (by capillary sequencing or MLPA) showed 100% concordance with NGS assay. Of the 310 CAH cases, biallelic mutations (pathogenic/ likely pathogenic variants involving both CYP21A2 gene copies) were detected in 256 (82.6%), heterozygous mutations in 13 (4.2 %), and none in 41 (13.2%). PCR products were purified and individual libraries were pooled and sequenced (Illumina). Orthogonal confirmation was performed by capillary sequencing (ABI 3500) and Multiplex Ligation-dependent Probe Amplification (MLPA, MRC-Holland). Locus-specific long-range PCR and gel electrophoresis of amplicons was followed by NGS where no visible 30 kb homozygous/whole gene deletion was observed. DNA was extracted from proband’s and parent’s(subset) blood. MethodsĬases with 21-OH CAH from 12 endocrine units across India were studied. Our objective was to develop an accurate NGS-based assay to characterize the mutation spectrum in CYP21A2 gene in Indian patients suspected to have 21-OH CAH. Next generation sequencing (NGS) provides data with high sequence coverage and depth. The conventional method for detection of mutations in the CYP21A2 gene is targeted capillary sequencing which is labor intensive and has limited multiplexing capability. Accurate diagnosis is required for management of Congenital adrenal hyperplasia (CAH). ![]()
0 Comments
Leave a Reply. |