DNA微阵列技术检测Duchenne型/Becker型肌营养不良患者的临床应用研究

Detection of Duchenne and Becker muscular dystrophy patients by DNA microarray

目的 研究检测Duchenne型肌营养不良 (DMD) /Becker型肌营养不良 (BMD)患者基因缺失的可行技术。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 30例DMD/BMD患者和 5例健康对照的基因进行检测分析。部分结果与PCR的方法作了比较。结果 应用简易DNA微阵列检测出 2 1例DMD/BMD患者具有不同程度的外显子缺失 ,10例经PCR检测得到了完全验证。结论 DNA微阵列技术检测DMD/BMD患者简便、准确、灵敏 ,可在临床诊断中应用。

Objective To explore the efficient method in detection of DMD/BMD patients.Methods 18 deletion-prone exon fragments of DMD gene were amplified via molecular cloning. They were used as probes and were spotted on the slides treated with APES and poly-lysine together by manual operation to make microarray. In addition, fragments of β-actin were used as positive contrast and those of pUC 19/EcoR I were used as negative. 30 DMD/BMD patients were detected for deletion in DMD gene with the microarray and 5 healthy people were done as normal control. Parts of the results were compared with PCR method.Results Different exon fragment deletion of DMD/BMD gene was detected in 21 patients by DNA microarray, and 10 of them were confirmed by PCR analysis.Conclusion DNA microarray assay is a convenient ,accurate and sensitive method in diagnosis of DMD/BMD patient.