Department of Human Genetics, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, The Netherlands.
Arch Med Res. 2012 May;43(4):312-6. doi: 10.1016/j.arcmed.2012.01.011. Epub 2012 Mar 3.
Next generation sequencing (NGS) approaches have revolutionized the identification of mutations underlying genetic disorders. This technology is particularly useful for the identification of mutations in known and new genes for conditions with extensive genetic heterogeneity. In the present study we investigated a consanguineous Pakistani family with intellectual disability (ID).
Genotyping was carried out using 250k and 6k SNP microarrays in order to perform homozygosity mapping and copy number variation (CNV) analysis. Targeted NGS was performed to identify the genetic defect in this family. qPCR was performed to validate and confirm the NGS result.
Homozygosity mapping positioned the causative defect on chromosome 2p25.3-p25.2. Subsequent targeted NGS revealed an intragenic deletion of five exons of the gene TPO.
NGS is a powerful method to uncover submicroscopic structural variations. This result demonstrates that an unbiased screening approach such as NGS can help to identify even unexpected disease-causing mutations.
下一代测序（NGS）方法的出现彻底改变了遗传疾病相关突变的鉴定。对于具有广泛遗传异质性的疾病，该技术特别有助于鉴定已知和新基因中的突变。在本研究中，我们调查了一个有智力障碍（ID）的巴基斯坦近亲家庭。
使用 250k 和 6k SNP 微阵列进行基因分型，以进行纯合性作图和拷贝数变异（CNV）分析。对该家族进行靶向 NGS 以鉴定遗传缺陷。进行 qPCR 以验证和确认 NGS 结果。
纯合性作图将致病缺陷定位在染色体 2p25.3-p25.2 上。随后的靶向 NGS 显示 TPO 基因的五个内含子发生了基因内缺失。
NGS 是一种揭示亚微观结构变异的强大方法。这一结果表明，像 NGS 这样的无偏筛选方法可以帮助识别甚至意想不到的致病突变。
