摘要
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入手，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 reractory 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。
Objeetive 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 gone 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. Results A missense mutation of GAT 〉CAT was identified at codon 1441 of the COL1A1 gone 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 gone mutation, D1441H, was identified in the Chinese pedigree with osteogenesis imperfecta type Ⅰ .
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
2009年第1期50-53,共4页
Chinese Journal of Medical Genetics
基金
南京军区医药卫生“十一五”科研基金（06MA136）
