用Transgreen/PI荧光复染法检测精子的存活率  

Viability assessment of human sperm using Transgreen and propidium iodide

作　　者：赵洪鑫[1] 袁瑶[2] 华敏敏[3] 张慧琴[2] 施惠娟[2] 

机构地区：[1]复旦大学,上海200032 [2]上海市计划生育科学研究所 [3]华东理工大学

出　　处：《中国男科学杂志》2008年第4期1-4,共4页Chinese Journal of Andrology

基　　金：国家自然科学基金项目(编号30571968);教育部留学回国人员科研启动基金;上海市浦江人才计划项目(06PJ14078)资助

摘　　要：目的探讨Transgreen/碘化丙锭(PI)荧光复染法用于检测人类精子存活率的可行性。方法以Invitrogen公司的Live/Dead Sperm Viability Kit的存活率结果为参考值,比较Transgreen/PI复染后精子的存活率结果与该试剂盒存活率结果的一致性;研究检测过程中试剂Transgreen和PI的加入顺序对存活率结果的影响;研究2h内Transgreen和PI染料对精子运动的影响。结果用Transgreen/PI复染法和Live/Dead Sperm Viability Kit复染法检测人类精子存活率,其结果无显著差异(P>0.05);精液中Transgreen和PI的加入顺序对精液存活率的检测结果没有影响(P>0.05);与空白对照相比,精液中加入Transgreen、PI及Transgreen/PI后2h内,精子的各项运动参数没有明显改变(P>0.05)。结论Transgreen/PI复染法与Live/Dead Sperm Vi-ability Kit一样,具有灵敏度高、重复性好及快速简便的优点,而且Transgreen/PI复染法价格低廉,可以作为检测精子存活率的常规实验室方法。Objective To establish a novel method with membrane-permeant DNA stain, Transgreen and propidium iodide （PI）, to evaluate the viability of sperm. Methods The assessment of human sperm viability using Transgreen/PI stainwas established; the results of Transgreen/PI and SYBR-14/PI which is commercially available as LIVE/ DEAD sperm viability kit were analyzed. The toxicity of Transgreen and PI respetively on sperm motility was observed. Results Based on data obtained at 0, 1, 2 and 24h after in vitro storage （37 ℃, 5% CO2）, there was no significant difference （P〉0.05） between Transgreen/PI and SYBR-14/PI to assess human sperm viability. There is no significant difference （P〉 0.05） between the results of assessment of sperm viability by different sequence of application of Transgreen and PI in stain. 2h after the application of Transgreen and PI, the stains have no prominent impact on motion parameters of sperm. Conclusion The dual association of Transgreen/PI procedure possesses good repeatability, high sensitivity, and real-time detection same as SYBR-14/PI. Besides, it is easily handled and much cheaper, 1% cost compared to SYBR- 14/PI. The combination of Transgreen and PI can be an effective tool for assessing the viability of human sperm in the future.

关 键 词：荧光染色 精子存活率 

分 类 号：R446.144[医药卫生—诊断学]