突变扩增系统法检测SRD5α2基因R227Q突变

Detection of R227Q mutation of SRD5α2 gene by amplification refractory mutation system

目的 建立一种简易、准确的2 型5α还原酶(SRD5α2)基因分析方法。方法 采用突变扩增系统,对23 例先天性尿道下裂患儿及正常标本进行SRD5α2 基因R227Q 突变分析,对阳性者进行DNA 测序证实。结果 23 例先天性尿道下裂患儿中,3 例为SRD5α2 基因R227Q 突变阳性(2 例为突变纯合子,1例为杂合子),与测序结果一致;其余及正常对照均为阴性。结论 此法简单、快速、准确、易于推广,可作为先天性尿道下裂中SRD5α2 基因突变的筛查方法。

Objective To set up a simple method for identifying the mutation of R227Q of SRD5α2 gene which is one of the most common mutations in congenital hypospadias in China.Methods The amplification refractory mutation system(ARMS) was employed in detecting the SRD5α2 gene R227Q mutation in congenital hypospadias confirmed by polymerase chain reaction single strand conformation polymorphism and DNA sequencing.Results The ARMS was successfully applied to the detection of the R227Q mutation of SRD5α2 gene.Three out of 23 congenital hypospadias were positive for R227Q mutation.The mutations determined by ARMS were in full agreement with those obtained by the DNA sequencing.Conclusion This is a simple,rapid and accurate method.It can be used for detecting the SRD5α2 gene R227Q mutation in congenital hypospadias and male pseudohermaphroditism.