Multiplex ligation-dependent probe amplification (MLPA) enhances the molecular diagnosis of aniridia and related disorders
Multiplex ligation-dependent probe amplification (MLPA) enhances the molecular diagnosis of aniridia and related disorders
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发表时间：
2008-05
期刊：
影响因子：
2.2
通讯作者：
E. Redeker;Annette S.H. de Visser;A. Bergen;M. Mannens
中科院分区：
文献类型：
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作者：
E. Redeker;Annette S.H. de Visser;A. Bergen;M. Mannens
PAX6基因的突变与无虹膜有关，无虹膜是一种先天性眼睛畸形，伴有严重的虹膜发育不良。然而，并不是所有的无虹膜病例都可以用PAX6基因的突变来解释。本研究的目的是利用多重连接依赖探针扩增技术(MLPA)提高无虹膜的分子诊断水平。从70例无虹膜先证者的外周血中提取总基因组DNA。进行聚合酶链式反应(PCR)，然后自动双向测序。此外，还进行了MLPA检查。测序后，我们在34名患者中发现了24个不同的PAX6基因点突变。在另外8名患者中，我们使用MLPA发现了PAX6基因的一个或多个外显子的缺失或PAX6基因的3‘调控区的缺失。这项工作证明了除了对PAX6基因的外显子进行测序外，还有必要在PAX6基因区域筛查较大的缺失。突变检测率将从49%提高到60%。这表明MLPA大大提高了无虹膜的分子诊断。
Mutations in the PAX6 gene have been implicated in aniridia, a congenital malformation of the eye with severe hypoplasia of the iris. However, not all aniridia cases can be explained by mutations in the PAX6 gene. The purpose of this study was to enhance the molecular diagnosis of aniridia using multiplex ligation-dependent probe amplification (MLPA). Total genomic DNA was isolated from peripheral blood of 70 unrelated probands affected with aniridia. Polymerase chain reaction (PCR) was performed followed by automated bidirectional sequencing. Additionally, MLPA was performed. We identified 24 different point mutations in the PAX6 gene in 34 patients after sequencing. In eight additional patients, we identified a deletion of one or more exons of the PAX6 gene or in the 3′ regulatory region of the PAX6 gene using MLPA. This work demonstrates the necessity to screen for larger deletions in the region of the PAX6 gene in addition to the sequencing of exons in the PAX6 gene. The mutation detection rate will increase from 49% to 60%. This shows that MLPA substantially enhances the molecular diagnosis of aniridia.