用双色荧光原位杂交技术检测罗伯逊易位携带者的精子染色体

目的 :探讨用双色荧光原位杂交技术 (D FISH)检测罗伯逊易位携带者的精子染色体的实验方法和应用价值。　方法 :采用荧光原位杂交 (FISH)技术 ,以Biotin标记的 13q 14 .3特异性探针和以Digoxigenin标记的 14 q11.1特异性探针对 2例罗伯逊易位携带者精子标本进行原位杂交 ,并统计精子间期核 13、14号染色体的杂交信号颗粒数量。　结果 :在显微镜下可见精子头部有以Biotin标记的 13q 14 .3特异性探针显示 1个绿色杂交信号 ,以Digoxi genin标记的 14 q11.1特异性探针显示 1个红色杂交信号 ,间期核背景经DAPI复染显示蓝色 ;共统计 30 0 0个精子间期核 ,显示 1个绿色 1个红色杂交信号的精子为 13q/ 14q ,占39.33% ;显示 1个绿色 2个红色杂交信号为 13q/ 14q ,14q ,占 11.5 7% ;仅显示 1个绿色杂交信号为 13q/ ,占 9.2 7% ;显示 2个绿色 1个红色杂交信号为 13q ,13q/ 14 q ,占12 .87% ;仅显示 1个红色杂交信号为 / 14 q ,占9.87% ;显示 2个绿色 2个红色杂交信号为 13q ,13q/ 14q ,14 q ,占12 .2 6 %。　结论 :用双色荧光原位杂交技术检测染色体结构异常患者的精子 ,可以分析其减数分裂过程中染色体分离规律 ,在人类生殖如显微授精和植入前胚胎遗传学诊断等方面具有非常广泛的应用价值

Detection of Sperm Chromosomes in Robertsonian Translocation Carriers by Dual-color Fluorescence in situ Hybridization

Objective: To study the technique of dual-color fluorescence in situ hybridization(D-FISH) and its application value in sperm chromosomes of Robertsonian translocation carriers. Methods: The technique of dual-color fluorescence in situ hybridization was used .Biotin labelled 13q 14.3 specific probe and Digoxigenin labeled 14q 11.1 specific probe were used for in situ hybridization of sperm specimens in 2 Robertsonian translocation carriers. Hybridization signals for chromosomes 13 and 14 in the sperm interphase nucleus were counted. Results: Under the microscope, Biotin labeled 13q 14.3 specific probe showed 1 green hybridization signal and Digoxigenin labeled 14q11.1 specific probe showed 1 red hybridization signal. Interphase nucleus counter-stained with DAPI showed blue. From the total of 3 000 sperm interphase nuclei, the positive rate for 1 green hybridization signal and 1 red hybridization signal was 13q/14q( 39.33%), for 1 green and 2 red was 13q/14q,14q( 11.57%), for 1 green was 13q/-( 9.27%), for 2 green and 1 red was 13q,13q/14q( 12.87%), for 1 red was -/14q( 9.87%)and for 2 green and 2 red was 13q,13q/14q,14q( 12.26%). Conclusions: D-FISH of the human sperm interphase nucleus can be applied to the determination of sperm chromosomes of Robertsonian translocation carriers and the analysis of the laws of chromosomal segregation in the meiosis. The technique can be widely used in such aspects of human reproduction as insemination under the microscope and preimplantation embryos genetic diagnosis.