Methylation-specific multiplex ligation-dependent probe amplification analysis of subjects with chromosome 15 abnormalities
Methylation-specific multiplex ligation-dependent probe amplification analysis of subjects with chromosome 15 abnormalities
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DOI：
10.1089/gte.2007.0061
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发表时间：
2007-12-01
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影响因子：
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通讯作者：
Butler, Merlin G.
中科院分区：
文献类型：
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作者：
Bittel, Douglas C.;Kibiryeva, Nataliya;Butler, Merlin G.
Prader-Willi综合征（PWS）和Angelman综合征（AS）是由15 q11-q13区域印记基因表达缺失引起的神经发育障碍。它们产生于该地区的类似缺陷，但起源不同。有两个公认的典型的15 q11-q13缺失，这取决于大小和几个诊断检测是可用的，但每个都有局限性。我们评估了由43个探针组成的甲基化特异性多重连接依赖性探针扩增（MLPA）试剂盒的有用性，以检测该区域的拷贝数变化和甲基化状态。我们使用MLPA试剂盒对82例15号染色体异常的受试者（62例PWS，10例AS和10例其他15号染色体异常的个体）和13例正常细胞遗传学结果进行基因分型。我们开发了一种MLPA探针分析算法，该算法正确识别了与PWS和AS相关的甲基化异常，并准确确定了先前分配的遗传亚型（包括印迹中心的微缺失）的拷贝数。此外，MLPA分析确定了远端15 q缺失和环15 s的拷贝数变化。MLPA是一种相对简单、成本效益高的技术，被发现对PWS和AS的甲基化状态、拷贝数和遗传亚型分析以及其他15号染色体异常有用且准确。
Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are neurodevelopmental disorders caused by loss of expression of imprinted genes from the 15q11-q13 region. They arise from similar defects in the region but differ in parent of origin. There are two recognized typical 15q11-q13 deletions depending on size and several diagnostic assays are available but each has limitations. We evaluated the usefulness of a methylation-specific multiplex ligation-dependent probe amplification (MLPA) kit consisting of 43 probes to detect copy number changes and methylation status in the region. We used the MLPA kit to genotype 82 subjects with chromosome 15 abnormalities (62 PWS, 10 AS and 10 individuals with other chromosome 15 abnormalities) and 13 with normal cytogenetic findings. We developed an algorithm for MLPA probe analysis which correctly identified methylation abnormalities associated with PWS and AS and accurately determined copy number in previously assigned genetic subtypes including microdeletions of the imprinting center. Furthermore, MLPA analysis identified copy number changes in those with distal 15q deletions and ring 15s. MLPA is a relatively simple, cost-effective technique found to be useful and accurate for methylation status, copy number and analysis of genetic subtype in PWS and AS, as well as other chromosome 15 abnormalities.