荧光原位杂交检测精子上游时间对XY精子比例的影响

Effect of Swim-up Time on Ratio of X-and Y-bearing Sperm

目的：运用荧光原位杂交（fluorescent in situ hybridization，FISH）技术检测精子改良上游过程中，不同上游时间对含X／Y精子的分离作用，探讨较长上游时间跨度（0～150分钟）对上游后XY精子比例的影响。方法：分析从志愿者取得的正常精液标本10份，每份均进行精子不同时间分批改良上游、固定和荧光原位杂交分析，计数分析X精子占XY精子总数的比例。结果：杂交区域内精子总杂交率为98．33％。未上游时X精子比例为平均为50．03％。通过SPSS方差分析：分时间段检测的X精子占XY精子总数的比例与未上游时差异无显著性（P〉0．05），不同组间差异无显著性（P均〉0．05）。结论：改良上游法对XY精子的分离没有显著效果，没有直接证据表明人工授精和体外受精．胚胎移植过程中精子的上游处理会影响到出生婴儿的性别比例。

Objective：To examine the separation of X- and Y-bearing spermatozoa in procedure of modified swimup by FISH（fluorescent in situ hybridization） ,and to find the effect of swim-up time on ratio of X- and Y-bearing spermatozoa. Methods： Use FISH to analyze the ratio of X- and Y-bearing spermatozoa after different swim-up time with X/Y dual-color CEP probes marked by FITC and Texas red. Results： 10 spermatozoa specimen from volunteers by masturbation were analyzed. A total ratio of hybridization is 98.33%. The ratio of X-bearing spermatozoa is 50.03% before swim-up. About 800 - 1000 spermatozoa were counted per slide. No significant difference of X-bearing and Y- bearing spermatozoa ratio between different swim-up time and before swim-up was found, among each individuals also,when analyzed by SPSS. Conclusions： No significant effect of separation of spermatozoa was found by modified swim-up method. There is no direct evidence to back up the opinion that swim-up method of spermatozoa used in IUI （intrauterine insemination） or IVF （in vitro fertilization） would led to unbalance of sex ratio after birth.