植入前遗传学诊断中卵裂球固定技术改进

An improved fixation technique of blastomeres for fluorescence in situ hybridization for preimplantation genetic diagnosis

目 的 改进植入前遗传学诊断卵裂球固定技术 ,减少卵裂球细胞丢失 ,获得更好的固定效率、更高的荧光原位杂交成功率 ,简化操作程序。方法 体外授精治疗周期废弃胚胎的 96个卵裂球细胞 ,分别用 3种不同的固定技术进行固定 ,固定后使用X、Y着丝粒探针进行荧光原位杂交 ,比较其固定效率和杂交效率。结果 改进方法固定率和杂交率均为10 0 %。而两种传统技术的固定率分别为 90 % ,83.3% ;杂交率为 80 % ,73.3%。结论 这种新的固定卵裂球技术从根本上消除了卵裂球标本的丢失 ,使PGD结果更为可靠 ;同时简化了操作程序 ,值得推广。

Objective:To improve existing preimplantation genetic diagnosis fixation techniques in order to eliminate the possibility of losing a cell during fixation and simplify the process of fixation of blastomere for preimplatation genetic diagnosis.Methods:Use three fixation techniques for 96 human blastomere of abandoned embryo in in vitro fertilization-embryo transfer cycle. Fluorescence in situ hybridization was accomplished after fixation using chromosome X and Y probes,and then analyzed fluorescent signals.Results:The improved fixation technique resulted in 100%fixation and 100% adequate FISH signals.Two conventional technique resulted in 90% and 83.3%fixation,and in adequate FISH signals in 80% and 73.3% respectively. Conclusion: This newly improved fixation technique essentially eliminates the possibility of losing a cell during fixation and simplifies the process of fixation of blastomere for preimplatation genetic diagnosis.