Preclinical validation of a microarray method for full molecular karyotyping of blastomeres in a 24-h protocol.
Preclinical validation of a microarray method for full molecular karyotyping of blastomeres in a 24-h protocol.
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DOI：
10.1093/humrep/dep452
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发表时间：
2010-04
期刊：
影响因子：
--
通讯作者：
Rabinowitz M
中科院分区：
文献类型：
--
作者：
Johnson DS;Gemelos G;Baner J;Ryan A;Cinnioglu C;Banjevic M;Ross R;Alper M;Barrett B;Frederick J;Potter D;Behr B;Rabinowitz M
植入前遗传学筛查（PGS）已被用于确定胚胎非整倍体。使用新的分子方法来确定胚胎核型的技术正在扩大PGS的范围。我们介绍了一种新的方法PGS，称为“父母的支持”，它利用从父母的DNA微阵列测量“清洁”单细胞微阵列测量胚胎细胞和明确计算每个拷贝数调用的信心。该方法区分有丝分裂和减数分裂拷贝错误，并确定非整倍性的亲本来源。用459个已知核型的单细胞验证表明，每个细胞的假阳性和假阴性率大致相当于“金标准”中期核型。大多数细胞与临床商业PGS服务并行运行。计算的置信度是保守的，大致一致的准确性。为了检测人类胚胎的倍性，该方法随后应用于26个分解的、冷冻保存的、卵裂期胚胎，总共134个单卵裂球。只有23.1%的胚为整倍体，而46.2%的胚为嵌合体整倍体。嵌合体影响57.7%的胚胎。有丝分裂和减数分裂错误的计数大致相等。母体减数分裂三体性占主导地位的父亲的三体性，和母体减数分裂三体性是负预测嵌合体整倍体胚胎。我们对PGS的新方法进行了主要的临床前验证，发现该技术的表现与中期核型大致相同。我们还直接测量了卵裂期人类胚胎的非整倍性机制，发现有丝分裂和减数分裂非整倍性的高比率和独特模式。
Preimplantation genetic screening (PGS) has been used in an attempt to determine embryonic aneuploidy. Techniques that use new molecular methods to determine the karyotype of an embryo are expanding the scope of PGS. We introduce a new method for PGS, termed ‘parental support’, which leverages microarray measurements from parental DNA to ‘clean’ single-cell microarray measurements on embryonic cells and explicitly computes confidence in each copy number call. The method distinguishes mitotic and meiotic copy errors and determines parental source of aneuploidy. Validation with 459 single cells of known karyotype indicated that per-cell false-positive and false-negative rates are roughly equivalent to the ‘gold standard’ metaphase karyotype. The majority of the cells were run in parallel with a clinical commercial PGS service. Computed confidences were conservative and roughly concordant with accuracy. To examine ploidy in human embryos, the method was then applied to 26 disaggregated, cryopreserved, cleavage-stage embryos for a total of 134 single blastomeres. Only 23.1% of the embryos were euploid, though 46.2% of embryos were mosaic euploid. Mosaicism affected 57.7% of the embryos. Counts of mitotic and meiotic errors were roughly equivalent. Maternal meiotic trisomy predominated over paternal trisomy, and maternal meiotic trisomies were negatively predictive of mosaic euploid embryos. We have performed a major preclinical validation of a new method for PGS and found that the technology performs approximately as well as a metaphase karyotype. We also directly measured the mechanism of aneuploidy in cleavage-stage human embryos and found high rates and distinct patterns of mitotic and meiotic aneuploidy.
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影响因子：
4.1
作者：
Frumkin, Tsvia;Malcov, Mira;Ben-Yosef, Dalit
通讯作者：
Ben-Yosef, Dalit
影响因子：
4
作者：
Hellani, Ali;Abu-Amero, Khaled;El-Akoum, Siham
通讯作者：
El-Akoum, Siham
影响因子：
6.1
作者：
Fragouli, E.;Wells, D.;Delhanty, J. D. A.
通讯作者：
Delhanty, J. D. A.
影响因子：
3
作者：
KALOUSEK, DK;HOWARDPEEBLES, PN;WILSON, RD
通讯作者：
WILSON, RD
影响因子：
1.7
作者：
Fragouli, E.;Wells, D.;Delhanty, J. D. A.
通讯作者：
Delhanty, J. D. A.