应用DNA微阵列技术检测缺失型DMD/BMD患者的研究

Detection deleted Duchenne and Becker muscular dystrophy patients by DNA microarray

目的研究Duchenne型肌营养不良(DMD)/Becker型肌营养不良(BMD)患者基因缺失检测的可行技术。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方法比较结果一致性。结果应用简易DNA微阵列检测出21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微阵列技术检测缺失型DMD/BMD患者简便、准确、灵敏,具有临床应用价值。

Objective： To explore the efficient method in gene deleted detection of DMD/BMD patients. Methods： 18 deletionprone 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. Differen, t exon fragment deletion of DMD 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 deleted DMD/BMD patient, and has practical significance in clinic.