Xie Yanxin, Xu Yanwen, Miao Benyu, Zeng Yanhong, Zhou Canquan
Center for Reproductive Medicine, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510080, P.R. China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2013 Jun;30(3):283-7. doi: 10.3760/cma.j.issn.1003-9406.2013.03.007.
To assess the value of array comparative genomic hybridization (array CGH) technique for preimplantation genetic diagnosis (PGD).
Array CGH was performed on three types of cells, which included 3-5 cells isolated from B2/C38/A1 embryonic stem cell lines, single cells isolated from two discarded normal fertilized embryos, and 10 blastocysts biopsied from 5 couples undergoing PGD for chromosomal translocations. For the 10 blastocysts, 8 were abnormal embryos, 1 appeared to be normal but showed arrested development, and 1 embryo was without any fluorescence signals. 24sure V3 or 24sure+ array chips were applied for CGH analysis. The results were analyzed with a BlueFuse Multi software.
(1) The results of cells from B2/C3/A1 embryo stem cells by array CGH were consistent with karyotyping analysis. (2) For the 6 single cell samples from two discarded embryos, 2 blastomeres from one embryo were diagnosed as with aneuploidy and a normal karyotype, respectively. Two out of 4 blastomeres biopsied from another embryo were normal, whilst the remaining two were diagnosed with aneuploidies of -22 and +13. Repeated detection with 24sure+ array was consistent with the 24sure V3 result. (3) Ten cell masses from 10 embryos in PGD cycles were successfully analyzed with array CGH, among which four were confirmed with fluorescence in situ hybridization (FISH) on day 3. In two of them, array CGH confirmed FISH diagnosis. For the remaining two, additional aneuploidies for chromosomes not tested by FISH were discovered by array CGH. Another embryo diagnosed as no signal by FISH was found to have trisomy 13 by array CGH. The remaining 5 embryos also showed discordant results by FISH and array CGH. One embryo from a Robertsonian translocation carrier was found to have monosomy 13 by FISH but trisomy 14 and additional aneuploidies by both 24sure V3 and 24sure+ chips. One embryo with many fragments and arrested development by D5 showed discordant results by FISH and array CGH. However, the FISH and array CGH results for other two embryos from this reciprocal translocation carrier were consistent. Three embryos with inconsistent results by FISH and array CGH had a chromosomal translocation involving q11 region.
Array CGH is useful for PGD applications for its capability to detect structural chromosomal abnormalities through screening of aneuploidies. However, the 24sure V3 array may not suit detection of translocations with breakpoints close to the q11 region of chromosomes.
评估阵列比较基因组杂交（array CGH）技术在植入前遗传学诊断（PGD）中的价值。
对阵列CGH技术在三种类型细胞上进行检测，这三种细胞分别为：从B2/C38/A1胚胎干细胞系分离出的3 - 5个细胞、从两个废弃的正常受精胚胎中分离出的单个细胞，以及从5对因染色体易位接受PGD的夫妇中获取的10个囊胚。对于这10个囊胚，8个为异常胚胎，1个看似正常但发育停滞，1个胚胎无任何荧光信号。应用24sure V3或24sure +阵列芯片进行CGH分析。结果用BlueFuse Multi软件进行分析。
（1）阵列CGH对B2/C3/A1胚胎干细胞的检测结果与核型分析一致。（2）对于来自两个废弃胚胎的6个单细胞样本，一个胚胎的2个卵裂球分别被诊断为非整倍体和正常核型。从另一个胚胎活检的4个卵裂球中，2个正常，其余2个被诊断为 - 22和 + 13的非整倍体。用24sure +阵列重复检测结果与24sure V3结果一致。（3）PGD周期中10个胚胎的10个细胞团成功进行了阵列CGH分析，其中4个在第3天通过荧光原位杂交（FISH）得到确认。其中2个，阵列CGH证实了FISH诊断。对于其余2个，阵列CGH发现了FISH未检测的染色体额外非整倍体。另一个FISH诊断为无信号的胚胎，阵列CGH检测发现有13三体。其余5个胚胎FISH和阵列CGH结果也不一致。一个来自罗伯逊易位携带者的胚胎，FISH检测为13单体，但24sure V3和24sure +芯片检测均显示有14三体及其他非整倍体。一个在第5天有许多碎片且发育停滞的胚胎，FISH和阵列CGH结果不一致。然而，来自这个相互易位携带者的另外两个胚胎FISH和阵列CGH结果一致。3个FISH和阵列CGH结果不一致的胚胎存在涉及q11区域的染色体易位。
阵列CGH通过筛查非整倍体来检测染色体结构异常的能力，使其在PGD应用中具有实用性。然而，24sure V3阵列可能不适用于检测断点靠近染色体q11区域的易位。
