[Establishment of an allele-specific PCR method for direct screening of CYP21A2 gene mutation].

[congenital adrenal hyperplasia]

To establish an allele-specific PCR method for detect screening of CYP 21 A 2 gene mutation .Allele-specific PCR primers and analogy primers were designed based on the sequence alignment of CYP 21 A 2 and CYP 21 AP genes . Genomic DNA was extracted from blood specimens of 4 patients with 21 -hydroxylase deficiency and 5 healthy controls and respectively amplified with allele-specific PCR primers and analogy primers and sequenced .Mutations of CYP 21 A 2 including IVS 2 -13 A /C >G , Arg 356 T rp and Arg 149 Pro were found with the established method in all of the 4 patients but not in the healthy controls . When detected with the analogy primers set , IVS 2 -13 A /C >G and Arg 356 Trp were observed in both patients and healthy controls .The allele-specific PCR-based method is a simple , effective and reliable method for the detection of CYP 21 A 2 gene mutation .