Fluorescence in situ hybridization analysis of two blastomeres from day 3 frozen-thawed embryos followed by analysis of the remaining embryo on day 5
Fluorescence in situ hybridization analysis of two blastomeres from day 3 frozen-thawed embryos followed by analysis of the remaining embryo on day 5
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DOI：
10.1093/humrep/deh094
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发表时间：
2004-03-01
影响因子：
6.1
通讯作者：
Martini, E
中科院分区：
文献类型：
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作者：
Baart, EB;Van Opstal, D;Martini, E
背景资料：在植入前遗传学诊断非整倍体筛查（PGD-AS）中，人类胚胎染色体嵌合现象可能导致假阳性或假阴性结果。因此，我们研究了从冻融胚胎的2细胞活检和荧光原位杂交（FISH）分析获得的结果是否代表了第5天剩余胚胎的染色体构成。方法：将冷冻保存的第3天胚胎解冻，并从存活的胚胎中活检两个卵裂球。对1、7、13、15、16、18、21、22、X和Y染色体进行FISH分析。活检后，将胚胎培养至第5天，并使用相同的探针板进行进一步分析。结果：总共有17个胚胎可用于诊断，基于第3天的两个卵裂球和第5天的验证性研究。在这17例病例中，有10例可以确认初步诊断。然而，只有6例细胞遗传学结果一致。除了10例诊断“正确”的病例外，还有6例假阳性结果和1例假阴性结果，均涉及马赛克现象。结论：研究两个卵裂球细胞核的染色体构成为研究早期胚胎发育中染色体嵌合现象的发生提供了一个很好的机会。第3天获得的结果的确认率取决于判读，从临床角度考虑时高于从细胞遗传学角度考虑时。
Background: Chromosomal mosaicism in human embryos may give rise to false positive or false negative results in preimplantation genetic diagnosis for aneuploidy screening (PGD-AS). Therefore, we have investigated whether the results obtained from a 2-cell biopsy of frozen-thawed embryos and fluorescence in situ hybridization (FISH) analysis are representative for the chromosome constitution of the remaining embryo on day 5. Methods: Cryopreserved day 3 embryos were thawed and from surviving embryos two blastomeres were biopsied. FISH analysis was performed for chromosomes 1, 7, 13, 15, 16, 18, 21, 22, X and Y. After biopsy, the embryos were cultured until day 5 and further analysed using the same probe panels. Results: In all, 17 embryos were available with a diagnosis based on two blastomeres on day 3 and confirmatory studies on day 5. In 10 of these 17 cases the initial diagnosis could be confirmed. However, in only six cases cytogenetic results were concordant. Besides the 10 cases with a 'correct' diagnosis, there were six false positive results and one false negative, all involving mosaicism. Conclusions: Investigating the chromosomal constitution of two blastomere nuclei offers a good opportunity to study the incidence of chromosomal mosaicism in early embryo development. The confirmation rate of the results obtained on day 3 depends on the interpretation and is higher when considered from a clinical than from a cytogenetic point of view.