Balasuriya A, Speyer B, Serhal P, Doshi A, Harper J C
UCL Centre for PG and D, Institute for Women's Health, University College London, UK.
Centre for Reproductive and Genetic Health, University College London Hospital, London, UK.
Reprod Biomed Online. 2011 May;22(5):428-36. doi: 10.1016/j.rbmo.2011.01.012. Epub 2011 Mar 11.
Sperm DNA damage is thought to be increased in men with male factor infertility. Previous studies suggest a correlation between sperm DNA fragmentation and aneuploidy. The sperm chromatin dispersion (SCD) test was modified to produce the Halosperm Kit. The SCD-fluorescent in-situ hybridization (FISH) test allows the simultaneous detection of DNA fragmentation and aneuploidy on the same sperm cell. The objectives of this study were to validate the SCD, SCD-FISH and Halosperm tests for the analysis of sperm DNA fragmentation and compare them to the sperm chromatin structure assay (SCSA). Semen samples from 20 males undergoing IVF/intracytoplasmic sperm injection were processed using FISH, SCD-FISH, SCD and Halosperm, and compared with SCSA results. There was a significant difference between FISH and SCD-FISH results in the detection of aneuploidy (P=0.000) and the level of sperm DNA fragmentation in the samples subjected to SCSA and SCD (P=0.001) or SCSA and SCD-FISH (P=0.001). There was no significant correlation between DNA fragmentation and aneuploidy. If sperm aneuploidy is to be determined, more reliable results will be obtained if FISH is performed rather than SCD-FISH. A lack of validation and unknown clinical significance question the value of DNA fragmentation assays. DNA damage in the male germ line may result in adverse clinical outcomes and the pathophysiology and clinical consequences of sperm DNA damage are being actively researched. Many DNA fragmentation assays such as the Halosperm Kit have been developed recently and are now available at a commercial level. Unfortunately, aimed at vulnerable couples with difficulty conceiving, many of these tests have not been clinically validated. Despite its plausible appeal and fervour of its supporters, the benefits of widespread DNA testing that only achieves the distressing of couples with the knowledge that effectual therapeutic strategies are absent are questionable. Commercially, however, it is no doubt lucrative. Analysis of gametes prior to the initiation of an IVF cycle may improve the quality of embryos transferred. The clinical and scientific community considers it a matter of urgency to translate the basic science behind how a cell prepares for fertilization into routine clinical practice. However, it is equally important, if not more, to allow the science behind such applications to draw level with its practice before its widespread implementation.
人们认为,男性因素不育患者的精子DNA损伤会增加。先前的研究表明精子DNA片段化与非整倍体之间存在关联。对精子染色质扩散(SCD)试验进行了改进,研发出了Halosperm试剂盒。SCD荧光原位杂交(FISH)试验能够在同一个精子细胞上同时检测DNA片段化和非整倍体。本研究的目的是验证SCD、SCD-FISH和Halosperm试验用于分析精子DNA片段化的效果,并将它们与精子染色质结构分析(SCSA)进行比较。对20名接受体外受精/卵胞浆内单精子注射的男性的精液样本进行FISH、SCD-FISH、SCD和Halosperm检测,并与SCSA结果进行比较。在检测非整倍体方面,FISH和SCD-FISH结果之间存在显著差异(P = 0.000),在接受SCSA和SCD(P = 0.001)或SCSA和SCD-FISH(P = 0.001)检测的样本中,精子DNA片段化水平也存在显著差异。DNA片段化与非整倍体之间没有显著相关性。如果要确定精子非整倍体,进行FISH检测比SCD-FISH能获得更可靠的结果。缺乏验证以及未知的临床意义对DNA片段化检测的价值提出了质疑。雄性生殖系中的DNA损伤可能导致不良临床结局,目前正在积极研究精子DNA损伤的病理生理学和临床后果。最近已经开发出许多DNA片段化检测方法,如Halosperm试剂盒,现在已有商业产品。不幸的是,针对难以受孕的脆弱夫妇,许多此类检测尚未经过临床验证。尽管其看似有吸引力且支持者热情高涨,但对于那些知道缺乏有效治疗策略却只能徒增痛苦的夫妇进行广泛的DNA检测,其益处值得怀疑。然而,在商业上,这无疑是有利可图的。在启动体外受精周期之前对配子进行分析可能会提高移植胚胎的质量。临床和科学界认为,将细胞如何为受精做准备的基础科学转化为常规临床实践是当务之急。然而,在广泛应用之前,让此类应用背后的科学与其实践相匹配同样重要,如果不是更重要的话。