【摘要】 背景:目前对Duchenne型肌营养不良(DMD)基因缺失的检测主要是Southern印迹或聚合酶链反应(PCR)的方法,在实际应用中有一定的局限性。DNA微阵列技术已广泛应用于基因突变的检测。目的:制备简易DNA微阵列检测DMD基因常见外显子缺失,作为一项新技术的方法学摸索,为开发更完善的DMD基因诊断芯片做准备。设计:非随机对照研究。地点和对象:5例患者来自2000-01/2001-12解放军第四军医大学西京医院神经内科门诊,所有患者均为男性。健康者为患者的父亲。方法:应用分子克隆的方法扩增DMD基因18个常见易缺失外显子片段,以此作为探针制备出简易DNA微阵列,对DMD患者和健康对照者的基因进行检测分析。主要观察指标:微阵列杂交结果;PCR结果。结果:应用简易DNA微阵列检测出4例DMD患者具有不同程度的外显子缺失,其结果与PCR验证相符,对照满意。结论:DNA微阵列技术适用于DMD基因缺失检测,具有简便、高通量、灵敏等特点。更多还原

【Abstract】 BACKGROUND:Recently, there are two main methods for the assay of Duchenne type muscular dystrophy(DMD):Southern blotting and polymerase chain reaction(PCR).While several limitations are found in the practice. DNA microarray has been widely used to test gene mutation. OBJECTIVE:To construct a simple DNA microarray for the measurement of the DMD gene common exon deletion,to grope for a methodology of new technique,and to prepare for the exploration of perfect DMD gene diagnosis chip. DESIGN:Non-randomized controlled study. SETTING and PARTICIPANTS:Five male patients were selected from the Out-patient Clinic of Neurology,Xijing Hospital of Fourth Military Medical University of Chinese PLA from January 2000 to December 2001.Men with physical fitness were the fathers of the patients. INTERVENTIONS:The 18 major deletion-prone exon fragments of DMD gene were amplified by the method of molecular cloning.These fragments were used as probes to prepare simple DNA microarray for the analysis of genes from DMD and healthy controlled patients. MAIN OUTCOME MEASURES:Results of microarray hybridization and PCR. RESULTS:Different degrees of exons deletion in 4 DMD patients were defected by simple DNA microarray. The results were in accordance with those of PCR verification. CONCLUSION:DNA microarray is suitable for the identification of Dystrophy gene deletion,with the features of convenience,high flux,sensitivity etc.更多还原