Establishment of an allele-specific PCR method for direct screening of CYP21A2 gene mutation.
- VernacularTitle:等位基因特异PCR结合测序检测CYP21A2基因突变的方法
- Author:
Haiqiang ZOU
1
,
2
;
Yan LIU
;
Weimin WANG
;
Fenghuan ZHANG
;
Baojian ZHAO
;
Junchao LIANG
Author Information
- Publication Type:Journal Article
- MeSH: Adrenal Hyperplasia, Congenital; enzymology; genetics; Alleles; Base Sequence; DNA Mutational Analysis; methods; DNA Primers; genetics; Humans; Molecular Sequence Data; Mutation; Polymerase Chain Reaction; methods; Steroid 21-Hydroxylase; genetics
- From: Chinese Journal of Medical Genetics 2014;31(4):479-482
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish an allele-specific PCR method for detect screening of CYP21A2 gene mutation.
METHODSAllele-specific PCR primers and analogy primers were designed based on the sequence alignment of CYP21A2 and CYP21AP 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.
RESULTSMutations of CYP21A2 including IVS2-13A/C>G, Arg356Trp and Arg149Pro 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, IVS2-13A/C>G and Arg356Trp were observed in both patients and healthy controls.
CONCLUSIONThe allele-specific PCR-based method is a simple, effective and reliable method for the detection of CYP21A2 gene mutation.
