Molecular analysis of SMA patients without homozygous SMN1 deletions using a new strategy for identification of SMN1 subtle mutations
Molecular analysis of SMA patients without homozygous SMN1 deletions using a new strategy for identification of SMN1 subtle mutations
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DOI：
10.1002/humu.20092
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发表时间：
2004-01-01
期刊：
影响因子：
3.9
通讯作者：
Cusin, V
中科院分区：
文献类型：
--
作者：
Clermont, O;Burlet, P;Cusin, V
脊髓性肌萎缩症是一种常见的常染色体隐性遗传病。在95%的患者中，SMA与5q13基因座相关联，在其中至少98%的患者中，发现SMN1纯合缺失。复合杂合型患者具有与微小突变相关的SMN1缺失，通过仅检测SMN1纯合子缺失的常见分子诊断测试，他们看起来没有缺失。在这些患者中，SMN1中的突变筛选因高度同源的SMN2基因的几个拷贝的存在而受到阻碍。在这里，我们提出了一种快速而可靠的策略，使用长程聚合酶链式反应检测SMN突变，避免了克隆和cDNA分析。使用这种方法，我们发现了10个突变，包括5个以前从未报道过的突变和5个反复发生的突变；其中一些可能是群体特有的。对这些突变中的5q13基因座的标记分析显示了共同的单倍型，支持共同祖先的假设，而不是热点序列。我们还评估了自动SSCA和DHPLC在突变扫描中的适用性。(C)2004年Wiley-Liss公司
Spinal muscular atrophy (SMA) is a common autosomal recessive disease. SMA is linked to the 5q13 locus in 95% of patients, and in at least 98% of them, the SMN1 homozygous deletion is found. Compound heterozygous patients, who have an SMN1 deletion associated with a subtle mutation, appear undeleted with the common molecular diagnostic test that detects only the homozygous absence of SMN1. In these patients, mutation screening in SMN1 is hampered by the presence of several copies of the highly homologous SMN2 gene. Here, we present a rapid and reliable strategy for detecting SMN mutations using long-range PCR, which avoids cloning and cDNA analysis. Using this method, we found 10 mutations, including five mutations never reported previously and five recurrent mutations; some of them are probably population-specific. Marker analysis of the 5q13 locus in these mutations showed common haplotypes, supporting the hypothesis of a common ancestor rather than a hot spot sequence. We also evaluate the suitability of automated SSCA and DHPLC for mutation scanning. (C) 2004 Wiley-Liss, Inc.