Pratap Harsha, Hottigoudar Suma Yekappa, Nichanahalli Kubera Siddappa, Chand Parkash
Department of Anatomy, All India Institute of Medical Sciences, New Delhi, India.
Department of Anatomy, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India.
J Pharmacol Pharmacother. 2017 Apr-Jun;8(2):45-49. doi: 10.4103/jpp.JPP_187_16.
The integrity of sperm deoxyribose nucleic acid (DNA) is one of the determinants that ensure normal fertilization, implantation, pregnancy, and the generation of a healthy progeny. Assessment of sperm DNA fragmentation has gained importance as a tool to provide significant information regarding sperm quality, and it can independently predict sperm fertilizing potential. The sperm chromatin dispersion (SCD) assay is one of the available techniques to detect sperm DNA damage with results comparable to the gold standard - Sperm Chromatin Structure Assay. We present here a detailed methodology of an alkaline modification of SCD that can be carried out with minimal laboratory equipment. The SCD assay is a modified halo assay that utilizes chemical methods to demonstrate sperm DNA fragmentation. It involves the embedding of sperms in an agarose medium followed by exposure to alkaline denaturation and deproteinization. The results are interpreted under a light microscope. A visual scoring system is utilized to differentiate the sperms with possible DNA fragmentation from those without fragmented DNA and to determine the sperm DNA fragmentation index (SDFI) % for each semen sample. The SDFI % is directly proportional to the quality of sperm. The SCD assay is a simple, cost-effective, and reliable technique that can detect sperm DNA fragmentation, thus providing information regarding sperm functional quality and reproductive capacity. It is of significance in clinical and research areas of andrology and reproductive medicine, toxicology, and pharmacotherapeutics.
精子脱氧核糖核酸（DNA）的完整性是确保正常受精、着床、妊娠以及生育健康后代的决定因素之一。评估精子DNA片段化作为一种提供有关精子质量重要信息的工具，其重要性日益凸显，并且它能够独立预测精子的受精潜力。精子染色质扩散（SCD）检测是可用于检测精子DNA损伤的技术之一，其结果与金标准——精子染色质结构检测相当。我们在此介绍一种SCD碱性改良方法的详细步骤，该方法使用最少的实验室设备即可进行。SCD检测是一种改良的晕环检测方法，利用化学方法来显示精子DNA片段化。它包括将精子包埋在琼脂糖培养基中，随后进行碱性变性和脱蛋白处理。结果在光学显微镜下进行解读。使用视觉评分系统来区分可能存在DNA片段化的精子与未发生DNA片段化的精子，并确定每个精液样本的精子DNA片段化指数（SDFI）%。SDFI%与精子质量直接相关。SCD检测是一种简单、经济高效且可靠的技术，能够检测精子DNA片段化，从而提供有关精子功能质量和生殖能力的信息。它在男科学和生殖医学、毒理学以及药物治疗学的临床和研究领域具有重要意义。
