双色荧光原位杂交检测男性不育患者精子X和Y染色体

目的建立男性不育患者精子核双色荧光原位杂交技术（FISH）的检测方法。方法将20例男性不育患者精子标本经低渗液处理、固定后制片，用二硫苏糖醇（DTT）使精子去凝集，变性后与双色荧光标记（CEPX、Y）探针杂交。结果20例患者的6672个精子中，X染色体精子率为47．78％，Y染色体精子率为47．24％，性染色体数目异常的精子率为4．53％，有效杂交率为99．55％。结论成功地建立了检测人精子X和Y染色体的检测方法。

The Establishment of an Approach to Dual-color Fluorescence in Situ Hybridization of Sperm Chromosome X and Y in Male Infertility

Objective We aim to establish an approach to dual-color fluorescence in situ hybridization （FISH） of sperm chromosome X and Y in male infertility. Methods Twenty cases＇samples were collected,later washed with 1 x PBS and treated with KCl hypotonic solution, then immobilized with methanol and acetic acid （3 ： 1 ） ,dropped on the slides and agglutinated with DTT. Finally,the dual-color FISH were performed on human sperm nucleus using centromeric probes for chromosome X and Y. Results There were 6 672 sperms were counted in this experiment,X chromosome rate Was 47.78%, Y chromosome rate was 47.24%, sex chromosome abnormal rate was 4.53%, and hybridization rate was 99.55%. Conclusion Dual-color FISH method to analyze sperm chromosome X and Y in male infertility had been set up successfully.