21-羟化酶缺陷症基因诊断方法的建立及应用  

Development and application of a method for molecular diagnosis of 21-hydroxylase deficiency

作　　者：马定远[1] 孙云[1] 陈玉林[1] 杨冰[1] 成建[1] 黄美莲[1] 张瑾[1] 张菁菁[1] 胡平[1] 林颖[1] 蒋涛[1] 许争峰[1] 

机构地区：[1]南京医科大学附属南京妇幼保健院,210004

出　　处：《中华医学遗传学杂志》2013年第1期49-54,共6页Chinese Journal of Medical Genetics

基　　金：江苏省医学创新团队与领军人才课题（LJ201109）;南京市科技发展计划项目（201201064）;南京医科大学科技发展基金重点项目（2010NJMUZl7）

摘　　要：目的建立21-羟化酶缺陷症的基因诊断方法并评价其临床应用价值。方法收集9例肾上腺皮质增生症患儿血样，采用全长基因直接测序法分析21-羟化酶编码基因CYP21A2的点突变，同时采用多重连接依赖探针扩增技术和位点特异性PCR-酶切多态分析大片段的CyP2IA2基因缺失或（和）转换突变。同时对患儿的双亲也进行了基因检测。结果9例患儿共发现点突变5种，分别为IVS2-13A／C〉G（9个等位基因）、P．Arg356Trp（1个等位基因）、ClusterE6（1个等位基因）、P．Gln318X（1个等位基因）和Promcony（1个等位基因）。除Promcony不能明确其功能外，其它4种均为致病突变。检测到的基因缺失或（和）转换突变有两种，一种为大片段缺失，导致CYP21A2基因单拷贝缺失，共发现3例患者（3个等位基因），另一种为CYP21A2基因重排后形成的CYP21AIP／CYP21A2嵌合基因，共发现3例患者（3个等位基因），并应用建立的基因诊断方案确定了全部患者的基因型。所有的突变均遗传自亲代。结论建立了21-羟化酶缺陷症的基因诊断方法。该方法不仅能特异性地检测点突变，而且能够检测大片段的缺失或（和）转换突变，因此具有较高的临床应用价值。Objective To develop a method for elucidating genetic basis of 21-hydroxylase deficiency. Methods 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. Results Nine children were analyzed. Point mutations of the C YP21A2 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 cony （1 allele）. While the former 4 mutations are pathogenic, the role of Prom cony mutation in the pathogenesis was uncertain. Three cases had entire CYP21A2 gene deletions （3 alleles）, three had CYP21AIP/CYP21A2 chimeric mutations （3 alleles）. The genotypes of all patients were determined. And all of the mutations were inherited from parents. Conclusion A rational method for detecting point mutations and large deletions/conversions of CYP21A2 gene has been established.

关 键 词：先天性肾上腺皮质增生症 21-羟化酶 突变 多重连接依赖探针扩增 

分 类 号：R440[医药卫生—诊断学]