多色荧光原位杂交检测人卵细胞染色体非整倍体方法的建立

Detection of aneuploidy in human oocyte by multicolor fluorescence in-situ hybridization.

目的 建立多色荧光原位杂交技术检测人卵细胞染色体非整倍体的方法。方法 取试管婴儿助孕技术后未能受精成功的卵细胞 ,于取卵后 1～ 3d固定 ,采用多色荧光原位杂交方法检测卵细胞 13,16 ,18,2 1和 2 2号染色体的情况。结果 正常未受精卵细胞中期染色体显示一个成对的杂交信号 ,每条染色单体显示一个单个信号 ;分裂相中多出或缺少一个成对杂交信号表明多余或缺少一条染色体 ;分裂相中多出或缺少一个单个信号表明多余或缺少一条染色单体 ;两个单个信号分离表明两条姐妹染色单体分离。结论 采用多色荧光原位杂交方法可以有效检测人卵细胞染色体非整倍体异常。

Objective: To establish the method of aneuploidy detection in human oocytes by multicolor fluorescence in-situ hybridization (M-FISH). Methods: The unfertilized oocytes were fixed 1-3 days after oocyte retrieval. M-FISH was performed according to the Vysis protocol to check the chromosome status in oocytes by using centromeric enumerator probes for chromosomes 16, 18 and locus-specific probes for chromosomes 13, 21 and 22. Results: A chromosome from normal human unfertilized oocytes shows a double dotted signal, while a chromatid shows a single dot signal. An additional or missing double dotted signal means the presence of an additional chromosome or absence of a chromosome in the metaphase. An additional or missing single dot signal means the presence of an additional chromatid or absence of a chromatid in the metaphase. The big distance between two single signals means the predivision of two sister chromatids. Conclusion: Multicolor FISH is a simple and efficient method to detect oocyte aneuploidy.