CAJ | 학술논문

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CYP21A2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)、Cluster E6(1个等位基因)、p.Gln318X(1个等位基因)和Prom cony(1个等位基因).除Prom conv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CYP21 A1 P/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.
목적 건립21-간화매결함증적기인진단방법병평개기림상응용개치.방법 수집9례신상선피질증생증환인혈양,채용전장기인직접측서법분석21-간화매편마기인CYP21A2적점돌변,동시채용다중련접의뢰탐침확증기술화위점특이성PCR-매절다태분석대편단적CYP21A2기인결실혹(화)전환돌변.동시대환인적쌍친야진행료기인검측.결과 9례환인공발현점돌변5충,분별위IVS2 13A/C＞G(9개등위기인)、p.Arg356Trp(1개등위기인)、Cluster E6(1개등위기인)、p.Gln318X(1개등위기인)화Prom cony(1개등위기인).제Prom conv불능명학기공능외,기타4충균위치병돌변.검측도적기인결실혹(화)전환돌변유량충,일충위대편단결실,도치CYP21A2기인단고패결실,공발현3례환자(3개등위기인),령일충위CYP21A2기인중배후형성적CYP21 A1 P/CYP21A2감합기인,공발현3례환자(3개등위기인),병응용건립적기인진단방안학정료전부환자적기인형.소유적돌변균유전자친대.결론 건립료21-간화매결함증적기인진단방법.해방법불부능특이성지검측점돌변,이차능구검측대편단적결실혹(화)전환돌변,인차구유교고적림상응용개치.
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 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 cony (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.Conclusion A rational method for detecting point mutations and large deletions/conversions of CYP21A2 gene has been established.