荧光原位杂交技术研究人类体外未受精卵的17号染色体非整倍体

目的 对比两种不同间期核制备方法的效果，并比较25－30岁和31－35岁这两个女性年龄组、不同的体外受精－胚胎移植（in vitro fertilization-embryo transfer,IVF-ET）指征、不同超排方案等与17号染色体非整倍体率之间的关系。方法 采用0.1% Tween 20/0.01 mol/L HCl和3：1的甲醇：冰醋酸两种方法制备人类未受精卵间期核，选用人类17号染色体端粒探针（17qter），按说明书步骤进行荧光原位杂交并在方法上稍作改进。结果 36个未受精卵中，正常17号单体24枚，二体7枚，三体5枚，非整倍体出现率为33.3%（12／36）；25－30岁和31－35岁这两个女性年龄组、不同IVF指征、不同超排方案的患者的17号染色体非整倍体发生率差异无显著性。结论 两种方法信号检出率均为100％，但前者间期核制备效果较好，易于操作，避免甲醇冰醋酸刺激性气味对环境的污染，卵母细胞17号染色体的非整倍性是造成体外受精失败的重要原因之一。

Fluorescence in situ hybridization study on the chromosome 17 aneuploidy of human oocytes failing to fertilize in vitro

OBJECTIVE: Compare two methods of preparing human unfertilized oocytes interphase nucleus and analyze the relationship between the different in vitro fertilization(IVF) indications, ovarian stimulation protocols, women's age and frequency of chromosome 17 aneuploidy. METHODS: Tarkowski's air-drying method(3:1 methanol:acetic acid) and Coonen's 0.1% Tween 20/0.01 mol/L HCl method were used to fix human unfertilized oocytes interphase nucleus, and telomeric probe of 17qter was used by standard fluorescence in situ hybridization(FISH) procedures to confirm chromosome 17qter aneuploidy. RESULTS: Of 36 human unfertilized oocytes, 24 were haploid (66.7%), 7 were disomic (19.44%), 5 were trisomic (13.89%). The overall frequency of aneuploidy was 33.3%. There were no differences between the protocols characterized by different maternal age, IVF indication, ovarian stimulation. CONCLUSION: Tarkowski's air dry method is as good as the method of Coonen's, but the latter method can avoid the smell pollution of the methanol and acetic acid, and it is easy to operate. The chromosome 17 aneuploidy is one of the factors to cause in vitro fertilization failure of human oocytes.