Nakajima T, Matsuo M, Nakamura H, Fujiwara Y
Department of Pediatrics, Kobe University School of Medicine.
Kobe J Med Sci. 1991 Feb;37(1):21-33.
We have analyzed 34 Japanese patients of 31 families with Duchenne muscular dystrophy (DMD) by Southern blot and PCR (polymerase chain reaction) to detect large deletions in the genomic dystrophin gene on the X chromosome. Fifteen families (48%) had various deletions in the dystrophin gene by Southern blot analysis using the dystrophin cDNA probes 1-2a, 2b-3 and 8 which effectively covers the high-risk regions of deletion. Lengths and positions of the gene deletions detected were variable. Although there were no common deletions of exons in the gene of the DMD patients examined with the above cDNA probes, the deletion frequency of each exon was almost the same as the result previously reported in Caucasian DMD patients. The PCR method was also applied to confirm gene deletions in 9 cases from the above 15 families by amplifying 6 different regions of the dystrophin gene. Eight out of 9 DMD cases had gene deletions at the same genomic regions as found by Southern blot analysis. In a single DMD case, the Southern blot analysis indicated the exon 12 deletion, but PCR successfully amplified a 331 bp DNA fragment containing exon 12 and flanking introns. This difference may arise by a large deletion except for the above small 331 bp sequence in the HindIII digests of dystrophin gene.
我们通过Southern印迹法和聚合酶链反应（PCR）分析了31个杜氏肌营养不良症（DMD）家族中的34名日本患者，以检测X染色体上基因组肌营养不良蛋白基因的大片段缺失。使用能有效覆盖缺失高风险区域的肌营养不良蛋白cDNA探针1-2a、2b-3和8进行Southern印迹分析，15个家族（48%）的肌营养不良蛋白基因存在各种缺失。检测到的基因缺失长度和位置各不相同。虽然用上述cDNA探针检测的DMD患者基因中没有外显子的共同缺失，但每个外显子的缺失频率与先前报道的白种人DMD患者的结果几乎相同。还应用PCR方法通过扩增肌营养不良蛋白基因的6个不同区域来确认上述15个家族中9例患者的基因缺失。9例DMD病例中有8例在与Southern印迹分析相同的基因组区域存在基因缺失。在1例DMD病例中，Southern印迹分析显示外显子12缺失，但PCR成功扩增出一个包含外显子12及其侧翼内含子的331 bp DNA片段。这种差异可能是由于肌营养不良蛋白基因的HindIII消化产物中除上述小的331 bp序列外存在大片段缺失所致。
