Laboratorio di Genetica Molecolare, IRCCS Oasi Maria SS, Troina (EN), Italy.
Exp Mol Med. 2010 Dec 31;42(12):842-8. doi: 10.3858/emm.2010.42.12.087.
Angelman syndrome (AS) is a severe neurobehavioural disorder caused by failure of expression of the maternal copy of the imprinted domain located on 15q11-q13. There are different mechanisms leading to AS: maternal microdeletion, uniparental disomy, defects in a putative imprinting centre, mutations of the E3 ubiquitin protein ligase (UBE3A) gene. However, some of suspected cases of AS are still scored negative to all the latter mutations. Recently, it has been shown that a proportion of negative cases bear large deletions overlapping one or more exons of the UBE3A gene. These deletions are difficult to detect by conventional gene-scanning methods due to the masking effect by the non-deleted allele. In this study, we have used for the first time multiplex ligation-dependent probe amplification (MLPA) and comparative multiplex dosage analysis (CMDA) to search for large deletions affecting the UBE3A gene. Using this approach, we identified a novel causative deletion involving exon 8 in an affected sibling. Based on our results, we propose the use of MLPA as a fast, accurate and inexpensive test to detect large deletions in the UBE3A gene in a small but significant percentage of AS patients.
天使综合征(AS)是一种严重的神经行为障碍,由位于 15q11-q13 上的印迹域的母源拷贝表达失败引起。有不同的机制导致 AS:母源微缺失、单亲二倍体、假定印迹中心缺陷、E3 泛素蛋白连接酶(UBE3A)基因突变。然而,一些疑似 AS 的病例仍然对所有这些突变均呈阴性。最近,已经表明一部分阴性病例携带重叠 UBE3A 基因一个或多个外显子的大片段缺失。由于非缺失等位基因的屏蔽作用,这些缺失很难通过常规基因扫描方法检测到。在本研究中,我们首次使用多重连接依赖性探针扩增(MLPA)和比较多重剂量分析(CMDA)来搜索影响 UBE3A 基因的大片段缺失。使用这种方法,我们在一个受影响的同胞中鉴定出一个涉及外显子 8 的新的致病缺失。基于我们的结果,我们建议使用 MLPA 作为一种快速、准确和廉价的检测方法,用于检测一小部分但具有显著意义的 AS 患者中 UBE3A 基因的大片段缺失。