The Sperm Chromatin Structure Assay (SCSA®) and other sperm DNA fragmentation tests for evaluation of sperm nuclear DNA integrity as related to fertility
The Sperm Chromatin Structure Assay (SCSA®) and other sperm DNA fragmentation tests for evaluation of sperm nuclear DNA integrity as related to fertility
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DOI：
10.1016/j.anireprosci.2016.01.017
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发表时间：
2016-06-01
影响因子：
2.2
通讯作者：
Evenson, Donald P.
中科院分区：
文献类型：
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作者：
Evenson, Donald P.
35年前，《科学》杂志发表了一篇关于精子DNA完整性与妊娠结局之间关系的开创性论文（240:1131），作为封面文章，展示了红色和绿色染色精子的荧光显微照片。流式细胞术数据显示，可育公牛和非可育公牛和男性的精子DNA完整性存在非常显著的差异。本研究利用热量（100℃，5分钟）对DNA链断裂部位的DNA变性，然后用吖啶橙（AO）染色，并测量5000个个体精子的绿色双链（ds） DNA和红色单链（ss） DNA荧光。随后，加热方案改为低pH方案，使DNA链断裂部位变性；热和酸的过程产生了相同的结果。SCSA数据是非常有利的双参数，具有1024通道（度）的红色和绿色荧光。数百篇关于在动物和人类中使用SCSA测试的出版物都证实了SCSA是一种非常有用的测试，可以确定雄性繁殖的可靠性。SCSA测试是一种快速，无偏差的流式细胞仪测量，提供具有卓越精度和可重复性的可靠统计数据。许多基因毒性实验显示了极好的剂量反应数据和极低的变异系数，进一步证实了SCSA是一种非常有效的精子DNA完整性检测方法。在SCSA检测被引入12年后，精子末端脱氧核苷酸转移酶介导的荧光素- dutp缺口末端标记（TUNEL）检测（1993年）被引入，作为精子DNA片段化的唯一另一种流式细胞术检测。然而，TUNEL测试也可以通过光学显微镜进行，但统计稳健性要低得多。COMET（1998年）和精子染色质弥散（SCD； HALO）（2003年）测试作为光学显微镜测试引入，不需要流式细胞仪。由于这些测试每个样本只能测量50-200个精子，因此它们缺乏流式细胞术测量的统计稳健性。只有SCSA测试具有固定协议的精确标准化。其他测试的许多变化使得比较男性因素不育风险的数据和阈值非常困难。这四种精子DNA片段化试验和光学显微镜吖啶橙试验（AOT）的数据有不同程度的相关性。(C) 2016年作者。Elsevier B.V.出版
Thirty-five years ago the pioneering paper in Science (240:1131) on the relationship between sperm DNA integrity and pregnancy outcome was featured as the cover issue showing a fluorescence photomicrograph of red and green stained sperm. The flow cytometry data showed a very significant difference in sperm DNA integrity between fertile and subfertile bulls and men. This study utilized heat (100 degrees C, 5 min) to denature DNA at sites of DNA strand breaks followed by staining with acridine orange (AO) and measurements of 5000 individual sperm of green double strand (ds) DNA and red single strand (ss) DNA fluorescence. Later, the heat protocol was changed to a low pH protocol to denature the DNA at sites of strand breaks; the heat and acid procedures produced the same results.SCSA data are very advantageously dual parameter with 1024 channels (degrees) of both red and green fluorescence. Hundreds of publications on the use of the SCSA test in animals and humans have validated the SCSA as a highly useful test for determining male breeding soundness. The SCSA test is a rapid, non-biased flow cytometer machine measurement providing robust statistical data with exceptional precision and repeatability. Many genotoxic experiments showed excellent dose response data with very low coefficient of variation that further validated the SCSA as being a highly powerful assay for sperm DNA integrity.Twelve years following the introduction of the SCSA test, the terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labelling (TUNEL) test (1993) for sperm was introduced as the only other flow cytometric assay for sperm DNA fragmentation. However, the TUNEL test can also be done by light microscopy with much less statistical robustness.The COMET (1998) and Sperm Chromatin Dispersion (SCD; HALO) (2003) tests were introduced as light microscope tests that don't require a flow cytometer. Since these tests measure only 50-200 sperm per sample, they suffer from the lack of the statistical robustness of flow cytometric measurements. Only the SCSA test has an exact standardization of a fixed protocol. The many variations of the other tests make it very difficult to compare data and thresholds for risk of male factor infertility. Data from these four sperm DNA fragmentation tests plus the light microscope acridine orange test (AOT) are correlated to various degrees. (C) 2016 The Authors. Published by Elsevier B.V.