Normand Elizabeth, Qdaisat Sadeem, Bi Weimin, Shaw Chad, Van den Veyver Ignatia, Beaudet Arthur, Breman Amy
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
Department of Obstetrics and Gynecology, Baylor College of Medicine, Houston, TX, USA.
Prenat Diagn. 2016 Sep;36(9):823-30. doi: 10.1002/pd.4866. Epub 2016 Jul 30.
Detection of genomic copy number abnormalities in a single cell using array comparative genomic hybridization (CGH) offers a promising non-invasive alternative for prenatal diagnosis. Our objective was to compare three commercially available whole-genome amplification (WGA) kits for their capacity to produce high quality DNA from single cells that is suitable for both molecular genotyping and array CGH.
We examined kit performance on unfixed, fixed and fixed/permeabilized lymphoblastoid cells. Molecular genotyping methods were used to evaluate the fidelity of amplified DNA for genomic profiling, while array CGH was used to assess copy number from single cells harboring trisomy 21, a DiGeorge syndrome deletion, a CMT1A duplication or a MECP2 duplication.
Molecular genotyping was achieved from single cells but performance varied between WGA kits. Furthermore, we consistently detected a dosage difference in sex chromosomes for gender mismatched hybridizations and for chromosome 21 in trisomy 21 cells. The 2.5 Mb DiGeorge syndrome deletion was also detected using all three WGA platforms, whereas the 1.3 Mb CMT1A and the 0.6 Mb MECP2 duplications were not consistently detected.
These data suggest that single cell molecular genotyping and copy number analysis can be accomplished when WGA conditions are optimized. © 2016 John Wiley & Sons, Ltd.
使用阵列比较基因组杂交（CGH）检测单细胞中的基因组拷贝数异常为产前诊断提供了一种有前景的非侵入性替代方法。我们的目的是比较三种市售的全基因组扩增（WGA）试剂盒从单细胞中产生高质量DNA的能力，这种DNA适用于分子基因分型和阵列CGH。
我们检测了试剂盒在未固定、固定和固定/通透化的淋巴母细胞上的性能。分子基因分型方法用于评估扩增DNA用于基因组分析的保真度，而阵列CGH用于评估携带21三体、迪格奥尔格综合征缺失、CMT1A重复或MECP2重复的单细胞的拷贝数。
从单细胞中实现了分子基因分型，但WGA试剂盒之间的性能有所不同。此外，我们始终在性别不匹配杂交的性染色体以及21三体细胞的21号染色体中检测到剂量差异。使用所有三个WGA平台也检测到了2.5 Mb的迪格奥尔格综合征缺失，而1.3 Mb的CMT1A和0.6 Mb的MECP2重复未被一致检测到。
这些数据表明，当WGA条件优化时，可以完成单细胞分子基因分型和拷贝数分析。© 2016约翰威立父子有限公司。
