[1]黄 诚,陆碧玉,代元平,等.精液样本保存方式对精子DNA 碎片率检测影响的实验研究[J].现代检验医学杂志,2023,38(03):180-183.[doi:10.3969/j.issn.1671-7414.2023.03.033]
　HUANG Cheng,LU Bi-yu,DAI Yuan-ping,et al.Experimental Study on the Effect of Semen Sample Preservation on Sperm DNA Fragmentation Index Test[J].Journal of Modern Laboratory Medicine,2023,38(03):180-183.[doi:10.3969/j.issn.1671-7414.2023.03.033]
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精液样本保存方式对精子DNA 碎片率检测影响的实验研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第38卷
期数:
2023年03期
页码:
180-183
栏目:
研究简报·实验技术
出版日期:
2023-05-15

文章信息/Info

Title:
Experimental Study on the Effect of Semen Sample Preservation on Sperm DNA Fragmentation Index Test
文章编号:
1671-7414（2023）03-180-04
作者:
黄 诚; 陆碧玉; 代元平; 黄李霜; 杨金铃; 黄丽华; 陈大宇; 蔡 稔; 严提珍
（柳州市妇幼保健院医学遗传科/ 柳州市出生缺陷预防与控制重点实验室，柳州市生殖医学重点实验室，广西柳州 545001）
Author(s):
HUANG Cheng; LU Bi-yu; DAI Yuan-ping; HUANG Li-shuang; YANG Jin-ling; HUANG Li-hua; CHEN Da-yu; CAI Ren; YAN Ti-zhen
（Department of Medical Genetics, Liuzhou Maternal and Child Health Hospital/ Liuzhou Key Laboratory of Birth Defect Prevention and Control / Liuzhou Key Laboratory of Reproductive Medicine, Guangxi Liuzhou 545001，China）
关键词:
精子DNA 碎片率;  流式细胞术;  样本保存
分类号:
R446.19
DOI:
10.3969/j.issn.1671-7414.2023.03.033
文献标志码:
A
摘要:
目的 研究精液样本新鲜度对精子DNA 碎片率（DNA fragmentation index，DFI）检测结果的影响，探讨在实际工作中该如何正确保存样本。方法 随机选择38 份2020 年12 月期间柳州市妇幼保健院生殖中心男科门诊日常送检的精液样本，采用流式细胞术（flow cyto metry，FCM）进行精子DFI 检测，每个样本充分液化后混匀分成4 等份，分为A，B，C 和D 共4 组：A 组当即检测，然后放置室温保存，每2h 再测1 次，共测得A1，A2，A3 三组数据；B 组放置2℃～ 8℃冰箱保存，每24h 检测1 次，连测3 天，共获得B1，B2，B3 三组数据；C 组放置-20℃冰箱冷冻，每周解冻测1 次，测完放回冰箱复冻，连测4 周，共获得C1，C2，C3，C4 四组数据；D 组放-20℃冰箱冷冻，30 天后解冻检测1 次，获得D1 组数据。将新鲜样本当即检测组(A1 组) 作为对照组，与其他组的DFI 结果进行单因素配对t检验，验证样本在不同保存方式下的结果变化。结果 A1 组DFI 较A2，C1，C2，C3，C4 和D 组[14.33（5.72 ～ 22.94）%vs 14.68（6.72 ～ 22.54）% ，14.59（6.52 ～ 22.66）%，14.73（6.62 ～ 22.95）%，14.91（6.89 ～ 22.93）%，14.96（7.20 ～ 22.72）%，14.65（6.85 ～ 22.45）%]， 差异无统计学意义（t=-0.827，-1.003，-1.483，-1.834，-1.786，-0.844， 均P>0.05）；A1 组DFI 低于A3，B1，B2，B3 组[14.33（5.72 ～ 22.94）% vs 16.44（8.07 ～ 24.81）%，20.99（11.93 ～ 30.05）%，20.71（11.19 ～ 30.23）%，23.10（12.57 ～ 33.63）%]，差异具有统计学意义（t=-3.091，-6.172，-5.108，-6.056，均P<0.05）。结论 为保障精子DFI 检测结果的可靠性，精液样本应在采集后2h 内完成检测，样本在室温或2 ～ 8℃放置时间过长会使检测结果偏高，若需较长时间保存建议及时将新鲜样本放置-20℃冰冻保存，时长可达一个月。
Abstract:
Objective To study the effect of semen sample freshness on the results of DNA fragmentation index (DFI), and explore how to properly preserve the samples in practical work. Methods 38 semen samples were randomly selected from the male outpatient department of the Reproductive Center of Liuzhou Maternal and Child Health Hospital during December 2020. Flow cyto metry（FCM） was used to detect the DFI of sperm. After each sample was fully liquefied, it was mixed and divided into four equal parts, and divided into four groups: group A was tested immediately, then stored at room temperature, and tested again every two hours for the data obtained from three groups of A1, A2 and A3. Group B was stored in a refrigerator at 2℃～ 8 ℃ , and tested once every 24 hours for 3 days to obtain data from three groups: B1, B2 and B3. Group C was frozen in the refrigerator at -20 ℃ , thawed and tested once a week. After the test, it was put back into the refrigerator for refreezing，and tested for 4 weeks to obtain data from four groups of C1, C2, C3 and C4. Group D was frozen in the refrigerator at -20 ℃ and thawed once after 30 days to obtain the data of group D1. The fresh sample immediate detection group (group A1) was taken as the control group, and the DFI results of other groups were tested by paired t-test to verify the changes in the results of samples under different storage methods. Results The DFI of group A1 was not significantly different from that of groups A2, C1, C2, C3, C4 and D1 [14.33（5.72 ～ 22.94）% vs 14.68（6.72 ～ 22.54）%, 14.59（6.52 ～ 22.66）%, 14.73（6.62 ～ 22.95）%， 14.91（6.89 ～ 22.93）%，14.96（7.20 ～ 22.72）%，14.65（6.85 ～ 22.45）%], and the differences were statistically significant（t=-0.827, -1.003, -1.483, -1.834, -1.786，-0.844, all P>0.05）. The DFI of group A1 was lower than that of groups A3, B1, B2 and B3[14.33（5.72 ～ 22.94）% vs 16.44（8.07 ～ 24.81）%，20.99（11.93 ～ 30.05）%，20.71 （11.19 ～ 30.23）%，23.10（12.57 ～ 33.63）%], and the differences were statistically significant （t=-3.091，-6.172，- 5.108，-6.056，all P<0.05）. Conclusion In order to ensure the reliability of the sperm DFI test results, the semen samples should be tested within 2h after collection. If the samples are stored at room temperature or 2 ～ 8℃ for too long, the test results will be high. If it needs to be stored for a long time, it is recommended to put the fresh samples at -20℃ for frozen storage, which can last for one month.

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备注/Memo

备注/Memo:
基金项目： 广西壮族自治区卫生和计划生育委员会科研课题（流式细胞术检测精子DNA 碎片率的室内质量控制研究（Z20180038）；精子过量残留细胞质对辅助生殖技术临床结局的相关性研究（Z20190165），非梗阻性无精子症的遗传因素分析及临床干预研究（Z20211191）；柳州市科技计划项目（2018AF10501）：柳州市医学遗传研究中心培养建设；广西医学高层次人才培养计划资助（G202003028）；柳州市个十百人才工程专项资金资助。
作者简介：黄诚（1986-），男，本科，副主任技师，研究方
更新日期/Last Update: 2023-05-15