Mutation screening of EXT1 and EXT2 by denaturing high-performance liquid chromatography, direct sequencing analysis, fluorescence in situ hybridization, and a new multiplex ligation-dependent probe amplification probe set in patients with multiple osteochondromas
Mutation screening of EXT1 and EXT2 by denaturing high-performance liquid chromatography, direct sequencing analysis, fluorescence in situ hybridization, and a new multiplex ligation-dependent probe amplification probe set in patients with multiple osteochondromas
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DOI：
10.2353/jmoldx.2008.070086
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发表时间：
2008-01-01
影响因子：
4.1
通讯作者：
Wuyts, Wim
中科院分区：
文献类型：
--
作者：
Jennes, Ivy;Entius, Mark M.;Wuyts, Wim
多发性骨软骨瘤（MO）是一种常染色体显性遗传的骨骼疾病，其特征是形成多个软骨覆盖的突起。MO是遗传异质性的，与EXT 1和EXT 2基因的突变有关。在这项研究中，我们描述了对63名临床和放射学诊断为MO的患者进行的广泛突变筛查。变性高效液相色谱分析显示43例患者的突变。通过荧光原位杂交和新开发的多重连接依赖性探针扩增探针组进行的额外缺失分析确定了1例基因内EXT 1易位患者，3例部分EXT 1缺失患者和1例部分EXT 2缺失患者。36名患者携带EXT 1突变（57%），12名患者携带EXT 2突变（19%）。我们表明，我们优化的变性高效液相色谱/测序/多重连接依赖性探针扩增方案代表了MO患者突变筛查的可靠且高度敏感的诊断策略。临床分析显示，在我们的MO患者队列中没有明确的基因型-表型相关性。
Multiple osteochondromas (MO) is an autosomal-dominant skeletal disorder characterized by the formation of multiple cartilage-capped protuberances. MO is genetically heterogeneous and is associated with mutations in the EXT1 and EXT2 genes. in this study we describe extensive mutation screening in a set of 63 patients with clinical and radiographical diagnosis of MO. Denaturing high-performance liquid chromatography analysis revealed mutations in 43 patients. Additional deletion analysis by fluorescence in situ hybridization and a newly developed multiplex ligation-dependent probe amplification probe set identified one patient with an intragenic EXT1 translocation, three patients with a partial EXT1 deletion, and one patient with a partial EXT2 deletion. Thirty-six patients harbored an EXT1 mutation (57%), and 12 had an EXT2 mutation (19%). We show that our optimized denaturing high-performance liquid chromatography/sequencing/multiplex ligation-dependent probe amplification protocol represents a reliable and highly sensitive diagnostic strategy for mutation screening in MO patients. Clinical analysis showed no clear genotype-phenotype correlation in our cohort of MO patients.