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在常规实验室操作中,镰状细胞贫血检测(SCD)试验和流式细胞术末端脱氧核苷酸转移酶介导的缺口末端标记法(TUNEL/FCM)能否互换使用来检测精子DNA损伤?

Can the SCD test and terminal uridine nick-end labeling by flow cytometry technique (TUNEL/FCM) be used interchangeably to measure sperm DNA damage in routine laboratory practice?

作者信息

Grèze Cécile, Guttmann Aline, Pons-Rejraji Hanae, Vasson Marie-Paule, Lornage Jacqueline, Ouchchane Lemlih, Brugnon Florence

机构信息

1Assistance Médicale à la Procréation, CECOS, Pôle Femme et Enfant, CHU Estaing, Clermont-Ferrand, France.

2Hôpitaux Universitaires de Strasbourg, Laboratoires de Biologie de la Reproduction et de Diagnostic préimplantatoire, Strasbourg, France.

出版信息

Basic Clin Androl. 2019 Dec 26;29:17. doi: 10.1186/s12610-019-0098-2. eCollection 2019.

Abstract

BACKGROUND

Numerous tests have been proposed to evaluate sperm DNA integrity. To assess the sperm chromatin dispersion (SCD) test in an andrology laboratory, twenty-five men attending Clermont-Ferrand (France) University Hospital's Center for Reproductive Medicine were recruited. Sperm DNA damage was measured in the same semen samples using the SCD test and the Terminal Uridine Nick-end Labeling by flow cytometry technique (TUNEL/FCM) after density gradient centrifugation.

RESULTS

SCD test reliability between readings, readers or slides was clearly established with very high agreement between measurements (Intraclass correlation coefficient (ICC) at 0.97, 0.95 and 0.98 respectively). Despite very good agreement between the SCD test and TUNEL/FCM (ICC at 0.94), the SCD test tended to slightly but significantly underestimate DNA damage compared with TUNEL ( = 0.0127). This systematic difference between the two techniques was - 3.39 ± 1.45% (mean ± SE).

CONCLUSIONS

Andrology laboratories using the SCD test to measure sperm DNA damage need to know that it appears to give slightly underestimated measurements compared to TUNEL/FCM. However, this systematic underestimation is very small in amplitude. Both techniques give almost perfectly congruent results. Our study underlines the importance for each laboratory to validate its method to assess sperm DNA damage before implementing it in routine andrology lab practice.

摘要

背景

已提出多种测试来评估精子DNA完整性。为了在一个男科实验室评估精子染色质扩散(SCD)测试,招募了25名前往法国克莱蒙费朗大学医院生殖医学中心就诊的男性。在密度梯度离心后,使用SCD测试和流式细胞术末端脱氧尿苷缺口末端标记技术(TUNEL/FCM)在相同的精液样本中测量精子DNA损伤。

结果

读数之间、读数者之间或载玻片之间的SCD测试可靠性已明确确立,测量之间具有非常高的一致性(组内相关系数(ICC)分别为0.97、0.95和0.98)。尽管SCD测试与TUNEL/FCM之间具有很好的一致性(ICC为0.94),但与TUNEL相比,SCD测试倾向于轻微但显著低估DNA损伤(P = 0.0127)。两种技术之间的这种系统差异为-3.39±1.45%(平均值±标准误)。

结论

使用SCD测试来测量精子DNA损伤的男科实验室需要知道,与TUNEL/FCM相比,它似乎会给出略低的测量值。然而,这种系统低估的幅度非常小。两种技术给出的结果几乎完全一致。我们的研究强调了每个实验室在将其评估精子DNA损伤的方法应用于常规男科实验室实践之前进行方法验证的重要性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/75f7/6933933/a28190fe360e/12610_2019_98_Fig1_HTML.jpg

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