CAJ | 학술논문

目的对Ⅰ型成骨不全(osteogenesis imperfecta,OI)1个家系进行分子诊断.方法从先证者的基因组DNA人手,自行设计30对引物,扩增产物涵盖全部COL1A1基因52个外显子及启动子区域,并以相应引物对PCR产物进行直接测序.针对突变位点,设计扩增阻滞突变系统(amplification refractory mutation system,ARMS)引物,在60名无关对照中进行突变筛查.结果在先证者的其中1条COL1A1等位基因上存在突变,即COL1A1基因第1441位(位于第52外显子,P30)发生了GAT→CAT改变.使原来编码的天冬氨酸被组氨酸取代(D1441H);其母亲的其中1条COL1A1等位基因上也存在相同突变,而正常对照相应的COL1A1基因序列与GenBank参考序列相同.ARMS分析显示,在60个无关对照中均未检测到D1441H突变.查阅国内外相关文献及COL1A1基因突变数据库,未发现有关COL1A1基因D1441H突变的报道.结论建立了基于COL1A1基因突变分析的成骨不全分子诊断方法 ,并在中国人Ⅰ型成骨不全患者中发现1个新的COL1A1基因D1441H突变.
목적 대Ⅰ형성골불전(osteogenesis imperfecta,OI)1개가계진행분자진단.방법 종선증자적기인조DNA인수,자행설계30대인물,확증산물함개전부COL1A1기인52개외현자급계동자구역,병이상응인물대PCR산물진행직접측서.침대돌변위점,설계확증조체돌변계통(amplification refractory mutation system,ARMS)인물,재60명무관대조중진행돌변사사.결과 재선증자적기중1조COL1A1등위기인상존재돌변,즉COL1A1기인제1441위(위우제52외현자,P30)발생료GAT→CAT개변.사원래편마적천동안산피조안산취대(D1441H);기모친적기중1조COL1A1등위기인상야존재상동돌변,이정상대조상응적COL1A1기인서렬여GenBank삼고서렬상동.ARMS분석현시,재60개무관대조중균미검측도D1441H돌변.사열국내외상관문헌급COL1A1기인돌변수거고,미발현유관COL1A1기인D1441H돌변적보도.결론 건립료기우COL1A1기인돌변분석적성골불전분자진단방법 ,병재중국인Ⅰ형성골불전환자중발현1개신적COL1A1기인D1441H돌변.
Objective To perform molecular diagnosis for a Chinese pedigree with osteogenesis imperfecta type Ⅰ.Methods Thirty pairs of primers were designed tO amplify all the 52 exons,exon boundaries and promoter region of the COL1A1 gene from genomic DNA of peripheral blood cells of the family members.The PCR products were purified and directly sequenced.To check the mutation in normal controls.the genomic DNA from peripheral blood cells of the index patient,his mother and 60 normal controls were analyzed by amplification refractory mutation system.ResultsA missense mutation of GAT ＞CAT was identified at codon 1441 of the COL1A1 gene from the family,which resulted in the replacement of aspartic acid by histidine(D1441H).This mutation was not found in a group of 60 normal controls.Conclusion The method for molecular diagnosis of osteogenesis imperfecta was established and a novel COL1A1 gene mutation,D1441H,was identified in the Chinese pedigree with osteogenesis imperfecta type Ⅰ.