Single cell segmental aneuploidy detection is compromised by S phase.
Single cell segmental aneuploidy detection is compromised by S phase.
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DOI：
10.1186/1755-8166-7-46
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发表时间：
2014
影响因子：
1.3
通讯作者：
Vermeesch JR
中科院分区：
文献类型：
--
作者：
Dimitriadou E;Van der Aa N;Cheng J;Voet T;Vermeesch JR
平衡易位携带者配子不平衡的风险很高，配子不平衡可能导致反复流产或出生缺陷。植入前遗传学诊断(PGD)通常用于选择平衡胚胎。这种选择目前主要是通过对卵裂球进行阵列CGH进行的。目前的方法没有考虑细胞周期的不同阶段，尽管在S期不同基因组区域的拷贝数状态是可变的。用来自3个不同染色体不平衡患者的细胞系评价单细胞阵列计算全息图的准确性。用流式细胞仪对不同的细胞周期时相进行分选，每个细胞系在每个细胞周期时相挑选10个单个细胞，利用最常用的PGD平台BAC阵列进行全基因组扩增和分析。与有效识别不平衡的G期相反，在感兴趣区域中，只有不到一半的探针指示17个S期细胞中存在像差，导致准确性降低。结果表明，S期细胞检测节段性染色体失衡的准确性降低，这可能是PGD误诊的原因之一。因此，被分析细胞的细胞周期时相非常重要，在分析过程中应该考虑到这一点。这一知识可能会指导未来的技术改进。
Carriers of balanced translocations are at high risk for unbalanced gametes which can result in recurrent miscarriages or birth defects. Preimplantation genetic diagnosis (PGD) is often offered to select balanced embryos. This selection is currently mainly performed by array CGH on blastomeres. Current methodology does not take into account the phase of the cell cycle, despite the variable copy number status of different genomic regions in S phase. Cell lines derived from 3 patients with different chromosomal imbalances were used to evaluate the accuracy of single cell array CGH. The different cell cycle phases were sorted by flow cytometry and 10 single cells were picked per cell line per cell cycle phase, whole genome amplified and analyzed by BAC arrays, the most commonly used platform for PGD purposes. In contrast to G phase, where the imbalances were efficiently identified, less than half of the probes in the regions of interest indicated the presence of the aberration in 17 S-phase cells, resulting in reduced accuracy. The results demonstrate that the accuracy to detect segmental chromosomal imbalances is reduced in S-phase cells, which could be a source of misdiagnosis in PGD. Hence, the cell cycle phase of the analyzed cell is of great importance and should be taken into account during the analysis. This knowledge may guide future technological improvements.
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DOI：
10.1093/humrep/der294
发表时间：
2011-11
期刊：
Human reproduction (Oxford, England)
影响因子：
--
作者：
Geraedts J;Montag M;Magli MC;Repping S;Handyside A;Staessen C;Harper J;Schmutzler A;Collins J;Goossens V;van der Ven H;Vesela K;Gianaroli L
通讯作者：
Gianaroli L
影响因子：
6.1
作者：
Alfarawati, S.;Fragouli, E.;Wells, D.
通讯作者：
Wells, D.
影响因子：
4
作者：
Campbell, Alison;Fishel, Simon;Thornton, Simon
通讯作者：
Thornton, Simon
影响因子：
2.6
作者：
Munne, Santiago
通讯作者：
Munne, Santiago
影响因子：
2.7
作者：
Manukjan, Georgi;Tauscher, Marcel;Steinemann, Doris
通讯作者：
Steinemann, Doris