【摘要】 目的建立适合于Duchenne型进行性肌营养不良(DMD)临床基因诊断的10对引物多重PCR方法 ,并探讨其临床应用价值。方法抽提55例DMD患者及正常对照者外周血DNA,选取肌营养不良蛋白基因的10对外显子引物,应用多重PCR反应方法 ,同时扩增10个特异性片段,扩增产物行聚丙烯酰胺凝胶电泳。结果 55例DMD患者中,28例(50.9%)存在外显子缺失,27例(49.1%)未检测到缺失;25例缺失片段集中于第45～53外显子,3例缺失片段集中于第12～19外显子。第50外显子缺失频率最高;其次为外显子45、47、51、52,再次之为外显子12、53。结论 DMD基因缺失片段主要分布于第45～53、12～19外显子2个缺失热点区域。该方法具有临床应用价值,是检测DMD基因缺失的首选方法 。更多还原

【Abstract】 Objective To establish and evaluate methods suitable for Duchenne muscular dystrophy (DMD)gene diagnosis by multiplex polymerase chain reaction (mPCR)using ten pairs of primers. Methods Genomic DNA of 55 patients with DMD was extracted and subjected to mPCR of the dystrophin genes by 10 pairs of primers. Results The optimized ten-primer-assembly of exons,reactive system and cycling condition of multiplex-PCR were important for the amplification with the clear bands of electrophoresis. Among the 55 patients with DMD,various exonic deletions were detected in 28 cases (50.9%).In twenty-five cases,the deletions were located in exon 45～53,and in three cases,the deletions were located in exons 12～19,The deletion of exon 50 ranked the first,that of exon 45,47,51 and 52 did the second,and that of exon 12 and 53 did the third. Conclusion DMD gene deletions are mainly distributed in two hot spots surrounding exons 45～53 and 12～19. This method is valuable for clinical diagnosis of DMD gene deletion.更多还原