双色FISH检测人精子染色体非整倍体方法的建立

Establishment of a Two-color Fluorescence in Situ Hybridization Method for the Detection of Aneuploid of Human Sperm Chromosomes

目的:建立用双色ＦＩＳＨ检测人精子染色体非整倍体的方法。方法:采用双色荧光原位杂交(ＦＩＳＨ)方法取适量精子标本用ＥＤＴＡ/ＰＢＳ处理,然后用二硫苏糖醇(ＤＴＴ),使精子去凝集。固定后滴片,然后与双色荧光直接标记探针杂交。结果;在ＯＬＹＭＰＵＳ荧光显微镜下可以清楚看到精子头部的蓝色杂交信号,头部有１个绿色荧光杂交信号的精子为Ｘ染色体精子(Ｘ精子),有１个红色荧光杂交信号的精子为Ｙ染色体精子(Ｙ精子)。精子头部有２个荧光杂交信号的精子为染色体数目异常精子。结论:双色荧光原位杂交(ＦＩＳＨ)方法可以用于测定人精子染色体非整倍体率的变化。

ve: To establish a method of two-color fluorescence in situ hybridization ( FISH ) for the detection of aneuploid of human sperm chromosomes. Methods: Sperm sample was washed with EDTA/PBS. The sperm heads were decondensed with dithiothreitol (DTT). Then, the sperm nuclei were hybridized with two - color fluorescence direct labeled enumeration probe, chromosome X α-satellite probe ( DXZ1 ) and chromosome Y-satellite probe ( DYZ3) . Results: Under an OLYMPUS fluorescence microscope, the hybridization signals in the sperm heads were clearly visible. The sperm with one green hybridization signal was X chromosome sperm (X sperm ) , and that with one red hybridization signal was Y chromosome sperm ( Y sperm ). Those with two hybridization signals in one sperm head should be abnormal in chromosome numbers. Conclusion The two-color FISH method can be used to detect the aneuploid frequency in chromosome of human sperms.