Development and application of a method for molecular diagnosis of 21-hydroxylase deficiency / 中华医学遗传学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To develop a method for elucidating genetic basis of 21-hydroxylase deficiency.</p><p><b>METHODS</b>Sanger sequencing of entire 21-hydroxylase coding gene CYP21A2 was carried out to detect point mutations, and multiplex ligation-dependent probe amplification (MLPA) and locus-specific PCR/enzyme restriction method were used to detect large deletions and conversion mutations.</p><p><b>RESULTS</b>Nine children were analyzed. Point mutations of the CYP21A2 gene have been identified as IVS2 13A/C>G (9 alleles), p.Arg356Trp (1 allele), Cluster E6 (1 allele), p.Gln318X (1 allele), and Prom conv (1 allele). While the former 4 mutations are pathogenic, the role of Prom conv mutation in the pathogenesis was uncertain. Three cases had entire CYP21A2 gene deletions (3 alleles), three had CYP21A1P/CYP21A2 chimeric mutations (3 alleles). The genotypes of all patients were determined. And all of the mutations were inherited from parents.</p><p><b>CONCLUSION</b>A rational method for detecting point mutations and large deletions/conversions of CYP21A2 gene has been established.</p>