CAJ | 학술논문

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.
배경:목전대Duchenne형기영양불량(DMD)기인결실적검측주요시Southern인적혹취합매련반응(PCR)적방법,재실제응용중유일정적국한성.DNA미진렬기술이엄범응용우기인돌변적검측.목적:제비간역DNA미진렬검측DMD기인상견외현자결실,작위일항신기술적방법학모색,위개발경완선적DMD기인진단심편주준비.설계:비수궤대조연구.지점화대상:5례환자래자2000-01/2001-12해방군제사군의대학서경의원신경내과문진,소유환자균위남성.건강자위환자적부친.방법:응용분자극륭적방법확증DMD기인18개상견역결실외현자편단,이차작위탐침제비출간역DNA미진렬,대DMD환자화건강대조자적기인진행검측분석.주요관찰지표:미진렬잡교결과;PCR결과.결과:응용간역DNA미진렬검측출4례DMD환자구유불동정도적외현자결실,기결과여PCR험증상부,대조만의.결론:DNA미진렬기술괄용우DMD기인결실검측,구유간편、고통량、령민등특점.
BACKGROUND:Recently, there are two main methods for the assay of Duchenne type muscular dystrophy(DMD): Southern blottingand 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 tech nique, 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 de fected by simple DNA microarray. The results were in accordance with those of PCR verification. CONCLUSION: DNA microarray is suitable for the identification of Dystro phy gene deletion, with the features of convenience, high flux, sensitivity etc.