Detailed analysis of 22q11.2 with a high density MLPA probe set
Detailed analysis of 22q11.2 with a high density MLPA probe set
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DOI：
10.1002/humu.20640
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发表时间：
2008-03-01
期刊：
影响因子：
3.9
通讯作者：
Emanuel, B. S.
中科院分区：
文献类型：
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作者：
Jalali, G. R.;Vorstman, J. A. S.;Emanuel, B. S.
染色体特异性低拷贝重复序列（LCR）的存在使22号染色体易于缺失和重复。目前用于检测22q11.2畸变的诊断程序是染色体分析结合荧光原位杂交（FISH）或基于PCR的多重连接依赖探针扩增（MLPA）。然而，在22q11.2中存在拷贝数变异（CNVs），其仅通过高分辨率平台如阵列比较基因组杂交（aCGH）检测到。我们报告了一个高清晰度MLPA（MLPA-HD）22 q11试剂盒，检测22号染色体长臂上37个位点的拷贝数变化的发展。这些包括通常在DiGeorge/velocardiofacial综合征（DGS/VCFS）中缺失的3-Mb区域，猫眼综合征（CES）区域，以及最近被证明缺失的22 q11中更远的区域。我们已经使用这种MLPA-HD探针组分析了363个先前良好的，具有22 q11处各种不同重排的特征的样品，并证明它可以以高灵敏度和特异性检测拷贝数改变。除了检测与DGS/VCFS相关的常见复发性缺失外，还检测到了22号染色体变异和新型畸变。这些包括该区域内的重复以及该区域远端的缺失。此外，MLPA-HD检测具有常见3-Mb缺失的患者之间的缺失终点差异。MLPA-HD试剂盒被提议作为具有22 q11畸变特征的个体的当前可用检测方法的具有成本效益的替代方案。在具有相关表型特征的患者中，该MLPA-HD探针组可替代FISH用于22q11.2缺失和重复的临床诊断。
The presence of chromosome-specific low-copy repeats (LCRs) predisposes chromosome 22 to deletions and duplications. The current diagnostic procedure for detecting aberrations at 22q11.2 is chromosomal analysis coupled with fluorescence in situ hybridization (FISH) or PCR-based multiplex ligation dependent probe amplification (MLPA). However, there are copy number variations (CNVs) in 22q11.2 that are only detected by high-resolution platforms such as array comparative genomic hybridization (aCGH). We report on development of a high-definition MLPA (MLPA-HD) 22q11 kit that detects copy number changes at 37 loci on the long arm of chromosome 22. These include the 3-Mb region commonly deleted in DiGeorge/velocardiofacial syndrome (DGS/VCFS), the cat eye syndrome (CES) region, and more distal regions in 22q11 that have recently been shown to be deleted. We have used this MLPA-HD probe set to analyze 363 previously well, characterized samples with a variety of different rearrangements at 22q11 and demonstrate that it can detect copy number alterations with high sensitivity and specificity. In addition to detection of the common recurrent deletions associated with DGS/VCFS, variant and novel chromosome 22 aberrations have been detected. These include duplications within as well as deletions distal to this region. Further, the MLPA-HD detects deletion endpoint differences between patients with the common 3-Mb deletion. The MLPA-HD kit is proposed as a cost effective alternative to the currently available detection methods for individuals with features of the 22q11 aberrations. In patients with the relevant phenotypic characteristics, this MLPA-HD probe set could replace FISH for the clinical diagnosis of 22q11.2 deletions and duplications.