Cuccaro Denis, Guarnaccia Maria, Iemmolo Rosario, D'Agata Velia, Cavallaro Sebastiano
1Institute of Neurological Sciences, National Research Council, Section of Catania, Catania, Italy; 2Section of Human Anatomy and Histology, Department of Biomedical and Biotechnological Sciences, University of Catania, Catania, Italy.
Curr Genomics. 2018 Sep;19(6):499-504. doi: 10.2174/1389202919666180122141425.
Copy Number Variants (CNVs) represent a prevailing type of structural variation (deletions or duplications) in the human genome. In the last few years, several studies have demonstrated that CNVs represent significant mutations in Alzheimer's Disease (AD) hereditability. Currently, innovative high-throughput platforms and bioinformatics algorithms are spreading to screening CNVs involved in different neurological diseases. In particular, the use of custom arrays, based on libraries of probes that can detect significant genomic regions, have greatly improved the resolution of targeted regions and the identification of chromosomal aberrations.
In this work, we report the use of NeuroArray, a custom CGH microarray useful to screening and further investigate the role of the recurring genomic aberrations in patients with confirmed or suspected AD.
The custom oligonucleotide aCGH design includes 641 genes and 9118 exons, linked to AD. The genomic DNA was isolated from blood samples of AD affected patients. The entire protocol of custom NeuroArray included digestion, labelling and hybridization steps as a standard aCGH assay.
The NeuroArray analysis revealed the presence of amplifications in several genes associated with AD. In the coding regions of these genes, 14,586 probes were designed with a 348 bp median probe spacing. The majority of targeted AD genes map on chromosomes 1 and 10. A significant aspect of the NeuroArray design is that 95% of the total exon targets is covered by at least one probe, a resolution higher than CGH array platforms commercially available.
By identifying with a high sensitivity the chromosomal abnormalities in a large panel of AD-related genes and other neurological diseases, the NeuroArray platform is a valid tool for clinical diagnosis.
拷贝数变异（CNV）是人类基因组中一种常见的结构变异类型（缺失或重复）。在过去几年中，多项研究表明CNV在阿尔茨海默病（AD）遗传易感性中代表着重要突变。目前，创新的高通量平台和生物信息学算法正在广泛应用于筛查与不同神经疾病相关的CNV。特别是，基于能够检测重要基因组区域的探针库的定制阵列的使用，极大地提高了靶向区域的分辨率和染色体畸变的识别能力。
在本研究中，我们报告了NeuroArray的使用情况，这是一种定制的比较基因组杂交（CGH）微阵列，有助于筛查并进一步研究确诊或疑似AD患者中反复出现的基因组畸变的作用。
定制的寡核苷酸aCGH设计包含641个基因和9118个外显子，与AD相关。从AD患者的血液样本中分离基因组DNA。定制NeuroArray的整个方案包括消化、标记和杂交步骤，作为标准的aCGH检测。
NeuroArray分析揭示了几个与AD相关的基因中存在扩增。在这些基因的编码区域，设计了14586个探针，中位探针间距为348 bp。大多数靶向AD基因位于1号和10号染色体上。NeuroArray设计的一个重要方面是，95%的总外显子靶点至少被一个探针覆盖，分辨率高于市售的CGH阵列平台。
通过高灵敏度地识别大量AD相关基因和其他神经疾病中的染色体异常，NeuroArray平台是临床诊断的有效工具。
