Darras B T, Koenig M, Kunkel L M, Francke U
Department of Human Genetics, Yale University School of Medicine, New Haven, CT 06510.
Am J Med Genet. 1988 Mar;29(3):713-26. doi: 10.1002/ajmg.1320290341.
DNA sequence polymorphisms (RFLPs) have been widely used as genetic markers for identification of the X chromosome that carries the mutation for Duchenne muscular dystrophy (DMD) in affected families, but serious limitations and pitfalls are associated with this approach [Darras et al., 1987]. The complementary DNA (cDNA) of the DMD gene has recently been isolated and shown to detect partial gene deletions in a large proportion of patients [Koenig et al., 1987]. Two prenatal studies are presented to illustrate how the unambiguous identification of deletion mutations by cDNA probes permits direct DNA-based diagnoses with high accuracy and in otherwise uninformative families. In a single proband family, DNA marker analysis had determined that the Xp21 chromosomal region present in the affected male was also carried by a male fetus in a subsequent pregnancy. Analysis of this family's DNA with probes covering the entire 14 kb cDNA revealed a small deletion in the affected male that was not present in the fetus nor in the mother. In the second family the fetus was a female deletion carrier identified by comparing intensities of restriction fragments. Since 1/3 of all DMD patients are thought to result from new mutations and most families have only single affected males, the cloned cDNA probes now available are likely to revolutionize DNA-based diagnostic studies in this disorder. More reliable, more rapid and less expensive than linkage studies with DNA polymorphisms, this method will be informative in the more than 50% of DMD/BMD cases that have deletion mutations.
DNA序列多态性(RFLPs)已被广泛用作遗传标记,用于在受影响的家族中鉴定携带杜氏肌营养不良症(DMD)突变的X染色体,但这种方法存在严重的局限性和缺陷[Darras等人,1987]。DMD基因的互补DNA(cDNA)最近已被分离出来,并显示在很大比例的患者中可检测到部分基因缺失[Koenig等人,1987]。本文介绍了两项产前研究,以说明cDNA探针如何明确鉴定缺失突变,从而在原本信息不足的家族中实现基于DNA的高精度直接诊断。在一个先证者家族中,DNA标记分析确定受影响男性中存在的Xp21染色体区域在随后的妊娠中也被一个男性胎儿携带。用覆盖整个14 kb cDNA的探针分析这个家族的DNA,发现受影响男性中有一个小的缺失,胎儿和母亲中均不存在。在第二个家族中,通过比较限制性片段的强度鉴定出胎儿是一个女性缺失携带者。由于所有DMD患者中有1/3被认为是新突变导致的,且大多数家族只有单个受影响男性,现有的克隆cDNA探针可能会彻底改变这种疾病基于DNA的诊断研究。这种方法比DNA多态性连锁研究更可靠、更快速且成本更低,对于超过50%有缺失突变的DMD/BMD病例将具有指导意义。