A modified multiplex ligation-dependent probe amplification method for the detection of 22q11.2 copy number variations in patients with congenital heart disease.
A modified multiplex ligation-dependent probe amplification method for the detection of 22q11.2 copy number variations in patients with congenital heart disease.
复制标题
一种改良的多重连接依赖性探针扩增方法，用于检测先天性心脏病患者的 22q11.2 拷贝数变异
DOI：
10.1186/s12864-015-1590-5
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发表时间：
2015-05-08
期刊：
影响因子：
4.4
通讯作者：
Sun K
中科院分区：
文献类型：
--
作者：
Zhang X;Xu Y;Liu D;Geng J;Chen S;Jiang Z;Fu Q;Sun K
背景：染色体22q11.2区域的拷贝数变异（CNVs）与一部分先天性心脏病（CHD）患者相关。准确、高效地检测CNV对CHD的遗传分析具有重要意义。本研究的目的是介绍一种新的方法命名为CNVplex®，一种高通量的分析技术，旨在有效地检测染色体CNVs，并探讨在22 q11.2位点的亚染色体不平衡的患病率与CHD患者从一个institute.Results：我们开发了一种新的技术，CNVplex®，用于高通量检测亚染色体拷贝数畸变。该方法在多重连接依赖探针扩增（MLPA）方法的基础上进行改进，引入了加长连接系统和4对通用引物，简化了探针的合成，显著提高了实验的灵活性。我们使用了110个样本，这是广泛的特点与染色体微阵列分析和MLPA，以验证新开发的方法的性能。同时，应用CNVplex®对上海儿童医学中心连续入选的818例CHD患者进行22q11.2位点亚染色体不平衡筛查。在方法学开发阶段，CNVplex®检测了先前用染色体微阵列分析和MLPA鉴定的所有拷贝数畸变，证明了100%的灵敏度和特异性。在验证阶段，CNVplex®分别在818例CHD患者中的39例和1例中检测到22q11.2缺失和22q11.2重复。我们的数据表明，22q11.2缺失的频率不同的冠心病患者的亚组。值得注意的是，22q11.2缺失在圆锥动脉干缺损（CTD）病例中比非CTD病例中更常见（P<0.001）。结论：CNVplex ®检测CNVs灵敏度高、特异性强，可替代MLPA用于已知基因组位点致病性CNVs的批量筛查。
Background:Copy number variations (CNVs) of chromosomal region 22q11.2 are associated with a subset of patients with congenital heart disease (CHD). Accurate and efficient detection of CNV is important for genetic analysis of CHD. The aim of the study was to introduce a novel approach named CNVplex®, a high-throughput analysis technique designed for efficient detection of chromosomal CNVs, and to explore the prevalence of sub-chromosomal imbalances in 22q11.2 loci in patients with CHD from a single institute.Results:We developed a novel technique, CNVplex®, for high-throughput detection of sub-chromosomal copy number aberrations. Modified from the multiplex ligation-dependent probe amplification (MLPA) method, it introduced a lengthening ligation system and four universal primer sets, which simplified the synthesis of probes and significantly improved the flexibility of the experiment. We used 110 samples, which were extensively characterized with chromosomal microarray analysis and MLPA, to validate the performance of the newly developed method. Furthermore, CNVplex® was used to screen for sub-chromosomal imbalances in 22q11.2 loci in 818 CHD patients consecutively enrolled from Shanghai Children's Medical Center. In the methodology development phase, CNVplex® detected all copy number aberrations that were previously identified with both chromosomal microarray analysis and MLPA, demonstrating 100% sensitivity and specificity. In the validation phase, 22q11.2 deletion and 22q11.2 duplication were detected in 39 and 1 of 818 patients with CHD by CNVplex®, respectively. Our data demonstrated that the frequency of 22q11.2 deletion varied among sub-groups of CHD patients. Notably, 22q11.2 deletion was more commonly observed in cases with conotruncal defect (CTD) than in cases with non-CTD (P<0.001). With higher resolution and more probes against selected chromosomal loci, CNVplex® also identified several individuals with small CNVs and alterations in other chromosomes.Conclusions:CNVplex® is sensitive and specific in its detection of CNVs, and it is an alternative to MLPA for batch screening of pathogenetic CNVs in known genomic loci.
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