精子染色质扩散试验及吖啶橙染色试验检测精子DNA完整性研究

Comparative study on determination of sperm DNA integrity by sperm chromatin dispersion test and by acridine orange staining test

目的探讨精子染色质扩散（sperm chromatin dispersion，SCD）试验及吖啶橙染色（AO）试验检测精子DNA完整性的应用价值。方法对32名已生育的成年男性健康对照者和27例特发性少精子症（idiopathic oligozoospermia，IO）患者同时进行SCD试验和AO试验，对检测结果进行分析。在SCD试验中，DNA完整性正常精子的DNA扩散形成大晕环或中晕环，而受损伤产生DNA碎片的精子不形成或形成很小的晕环。用AO试验检测，正常精子DNA为双链，染成绿色，不成熟或损伤精子DNA为单链，染成红色、橙色或黄色。结果SCI）试验检测IO患者的大晕环、中晕环、小晕环和无晕环精子百分率分别为（49．9±13．8）％、（11．5±5．4）％、（11．9±6．1）％和（26．7±10．0）％，健康对照组分别为（73．2±6．2）％、（14．7±6．3）％、（6．8±2．9）％及（5．3±2．2）％，2组比较，差异有统计学意义（t=8．576，P〈0．01；t=2．083，P〈0．05；f=4．284，P〈0．01；t=11．823，P〈0．01）；IO患者的DNA损伤精子的百分率为（38．6±12．1）％，健康对照组为（12．1±5．2）％，2组比较差异有统计学意义（t=11．995，P〈0．01）。AO试验检测10患者的单链DNA精子百分率为（45．5±13．8）％，健康对照组为（39．8±13．3）％，差异无统计学意义（t=1．626，P〉0．05）。结论精子DNA完整性异常可导致男性不育。SCD试验是一种有效的精子DNA完整性检测方法。

Objective To analyze and compare the accuracy and utility of determining sperm DNA integrity by sperm chromatin dispersion （SCD） test and acridine orange staining （AO） test. Methods The level of DNA fragmentation was determined by SCD test and AO test in 32 adult healthy fertile men and 27 idiopathic oligozoospermia （IO） patients. Sperm nuclei with large DNA dispersion halos or with mediumsized halos were normal and nuclei with small-sized halos or no halo were abnormal. The normal sperm DNA was double strands and stained by AO as green. The damaged sperm DNA was single strand, which were stained as green for native DNA and red for denatured DNA. Results The percentage of sperm nuclei with large halos, medium-sized halos, small-sized halos and no halo evaluated by SCD test in IO patients were （49.9±13.8）%, （11.5 ±5.4）%, （11.9 ±6. 1）% and （26.7±10.0）%,and they were （73.2± 6. 2 ） %, （ 14. 7 ± 6. 3 ） %, （ 6. 8 ± 2. 9 ） % and （ 5.3± 2. 2 ） % in healthy control group, respectively. There was significant difference between two groups （ t = 8. 576, P 〈 0. 01 ; t = 2. 083, P 〈 0. 05 ; t = 4. 284, P 〈 0. 01 ; t = 11. 823,P 〈 0. 01 ）. The percentage of sperm DNA fragmentation was （ 38.6 ± 12. 1 ） % in I0 patients and was （12. 1 ± 5.2）% in fertile men, respectively. A statistically significant difference was found between IO patients and healthy control group under SCD test （t = 11. 995,P 〈 0. 01 ）. AO test showed no significant differences between IO patients and healthy control group （t = 1. 626 ,P 〉0. 05）. The percentage of sperm DNA fragmentation evaluated by AO test in IO patients was （45.5 ± 13.8）% ,and it was （39. 8 ± 13.3 ）% in healthy control group. Conelnsions Sperm DNA fragmentation may lead to male infertility. The SCD is effective in testing the sperm DNA fragmentation as a screening procedure to determine semen quality during basic infertility investigation for clinical use.