变性梯度凝胶电泳在遗传性多发性外生性骨疣基因诊断中的应用

目的:采用检测遗传性多发性外生性骨疣(hereditary multiple exostoses,HME)患者EXT2基因的突变,分析该方法的敏感性及用于HME基因诊断的可行性.方法:收集5个HME家系和3个HME散发患者,利用变性梯度凝胶电泳方法对EXT2基因的所有编码外显子及其旁侧内含子序列进行检测,并对出现异常构象的片段进行DNA测序分析.结果:在两个家系中分别发现了一种 A313T的无义突变和一种319insGT的移码突变.结论:在HME患者中发现了2个EXT2基因致病突变,DGGE可作为基因诊断理想的候选方法.

Denaturant gradient gel electrophoresis in the genetic diagnosisof hereditary multiple exostoses

ObjectiveTo detect the mutations of EXT2 gene in hereditary multiple exostoses (HME) families and to investigate the sensitivity of denaturant gradient gel electrophoresis (DGGE) in screening the mutations in EXT2 gene. MethodsFive HME families and 3 sporadic patients were screened for the mutation detection in all exons of EXT2 gene covering the coding sequence and the flanking intronic sequence by DGGE, and DNA sequencing was performed for products with abnomal conformation. ResultsAmong these HME patients, we found 2 disease-causing mutations: A313T (nonsense mutation) and 319insGT (frameshift mutation). ConclusionTwo mutations of EXT2 gene are identified in the sample. DGGE can be an ideal choice for gene diagnoses of HME.