用于精子DNA碎片指数检测的精液标本处理方式探讨  

Investigating the method of semen treatment for the detection of sperm DNA fragmentation index

作　　者：韦庭侣 汪彩珠[1] 孙克俭 吴柱连 潘彦文 甘贤优[1] 周红[1] 舒金辉[1] Wei Tinglyu;Wang Caizhu;Sun Kejian;Wu Zhulian;Pan Yanwen;Gan Xianyou;Zhou Hong;Shu Jinhui(Reproductive Medical Center,Maternity and Child Health Hospital of Guangxi Zhuang Autonomous Region,Nanning 530000,China)

机构地区：[1]广西壮族自治区妇幼保健院生殖医学中心,南宁530000

出　　处：《中华生殖与避孕杂志》2023年第12期1260-1263,共4页Chinese Journal of Reproduction and Contraception

基　　金：广西自然科学基金(2018JJB14012);广西卫生健康委员会自筹经费课题(Z20200627, Z20210016)。

摘　　要：目的探讨不同的精液处理方式对精子DNA碎片指数(DNA fragmentation index, DFI)检测结果的影响, 评价当前精液处理方式对男性生育力评估的准确性。方法本研究采用自身配对设计, 选取2021年8月至2022年1月期间在广西壮族自治区妇幼保健院生殖医学中心行体外受精助孕患者在取卵当日获得的50份精液。每一份精液分为3组, 进行不同方法处理, A组:精液样本混匀后留取精液原液50 μL;B组:密度梯度离心后留取精液与梯度液交界层的精液50 μL;C组:密度梯度离心后, 吸取精子沉淀团加入含有1.0 mL精子洗涤液的试管底部置于培养箱内上游15 min后, 吸取50 μL上游液。精液处理后, 进行精液常规检测和精子DFI检测。采用Pearson进行分析DFI和精子不动精子百分率、精子前向运动百分率的相关性。结果 C组的精子活动率[(96.83±2.28)%]显著高于A组[(57.16±11.28)%]和B组[(22.54±9.35)%], 组间两两比较和3组之间比较差异均有统计学意义(均P<0.001)。B组的不动精子百分率[(77.46±9.35)%]、DFI[37.18%(30.41%, 47.80%)]高于C组[(3.14±2.31)%、0.78%(0.00%, 2.07%)]和A组[(42.83±11.28)%、22.00%(14.75%, 29.25%)], 组间两两比较和3组之间比较差异均有统计学意义(均P<0.001)。Pearson分析结果显示, A组和B组的DFI与不动精子百分率呈正相关(r=0.304, P=0.032;r=0.612, P<0.001), B组DFI与精子前向运动百分率呈负相关(r=-0.517, P<0.001)。结论对于同一份精液, 不动精子的DFI显著高于活动精子的DFI。精液原液的DFI值因受不动精子的干扰, 不能真实反映参与受精的活动精子的DNA情况, 提示应该选择梯度离心法、上游法或其他方法收集的活动精子进行DFI检测, 这样更能准确地评估男性生育力。Objective To investigate the effects of different semen sample collection methods on sperm DNA fragmentation index(DFI)test results,then to evaluate the accuracy of the current semen sample collection method in the assessment of male fertility.Methods In this study,50 semen sample obtained on the day of oocyte retrieval from patients undergoing in vitro fertilization at the Reproductive Medical Center in Maternity and Child Health Hospital of Guangxi Zhuang Autonomous Region from August 2021 to January 2022 were collected.For each semen,a small amount of samples were collected in three different retention methods for routine semen and sperm DFI testing.Three different ways of retaining samples were as follows:group A,after mixing of the semen,50μL sample was directly collected;group B,after density gradient centrifugation,50μL sample was collected at the interface between semen and gradient fluid;group C,after density gradient centrifugation,the sperm pellet was upstream,then 50μL sample was collected from upstream liquid.After semen treatment,routine semen testing and sperm DFI testing were performed.Pearson was used to analyze the correlation between DFI and the percentage of immobile sperm and the percentage of forward sperm movement.Results The sperm motility rate of group C[(96.83±2.28)%]was significantly higher than that of group A[(57.16±11.28)%,P<0.001]and group B[(22.54±9.35)%,P<0.001],and there was a statistical difference among the three groups.The immotile sperm rate of group B sample was(77.46±9.35)%,which was significantly higher than that of samples from group C[(3.14±2.31)%,P<0.001]and group A[(42.83±11.28)%,P<0.001].There was also a statistical difference in DFI among the three groups(P<0.001).The DFI of group B[37.18%(30.41%,47.80%)]was significantly higher than that of group A[22.00%(14.75%,29.25%),P<0.001]and group C[0.78%(0.00%,2.07%),P<0.001].Pearson analysis results showed that the DFI of group A and group B was positively correlated with the percentage of immobile sperm(r=0.304,P=

关 键 词：DNA碎片指数 精子活力 不动精子 精液处理方法 

分 类 号：R714.8[医药卫生—妇产科学]