Although a specific upper or lower limit for predicting pregnancy is still under debate for major dna integrity parameters eg. Vials must be labelled with name, dob and date of sample. Intriguingly, cerium oxide ceo2 nanoparticles can manifest either anti or prooxidant activity as a function of differential ph of various subcellular localizations. Sperm dna sperm dna is organized in a specific manner that keeps the unclear chromatin compact and stable. Dna fragmentation, and formation of membrane bound vesicles apoptotic bodies apoptosis is both a very complex and very important process and dysregulations in the apoptosis machinery may lead to very severe diseases. In this study, the dna fragmentation index and the degree of sperm decondensation were measured using the sperm chromatin structure assay before and after. The clinical utility of sperm dna integrity testing. Some thoughts on the utility of sperm nuclear dna fragmentation assays in reproduction. Sperm dna fragmentation sdf is also called sperm dna damage. It can be done intentionally by laboratory personnel or by cells, or can occur spontaneously. Male factor accounts for approximately 20% of cases of male infertility.
There is significant relationship observed between sperm aneuploidy and dna fragmentation. However, dna fragmentation may affect the clinical pregnancy rate oleszczuk et al. Dna fragmentation may affect the fertilization rate after in vitro fertilization ivf. Diagnostic value of sperm dna fragmentation and sperm high. The only way to evaluate male fertility was actually limited to sperm analysis, which can provide information about 3 aspects. Dfi, known as sperm dna fragmentation index, was established to evaluate sperm chromatin integrity, and has gained increasing application for its diagnostic capabilities of male fertility potential and pregnancy outcome 2, 3. In a smaller danish study including 48 couples, no pregnancies were observed in couples, where the male dna fragmentation exceeded 27% boehansen et al. This threshold value for sperm dna fragmentation in our study differs from the value suggested by the test manufacturer, which established a fragmentation index of 30%. Our results indicated that pfc contamination was present in 58% of subjects included in the study.
The effect of sperm dna fragmentation index on the outcome of. The clinical threshold is a dna fragmentation index dfi of 30% that means 30% of the sperm have damage with quantification into moderate or high damage and 70% have no detectable damage. A growing body of evidence suggests that resistance to apoptosis is a feature. Total reactive antioxidant potential and dna fragmentation. The sureselect enzymatic fragmentation kit produces similar dna fragmentation patterns from dna samples with various quality. Human sperm nuclear dna fragmentation assays and their. Chromosomal aneuploidies and dna fragmentation of human. What should be done for men with sperm dna fragmentation.
This loss of spatial information requires an intensive computational process to reassemble the original dna sequence. However in some specific cases we could not come across with any possible etiology. Sperm dna fragmentation sdf testing measures the quality of sperm as a dna package carrier, and it therefore is more significant than the parameters analyzed in previous semen analyses. Highly similar dna fragmentation profiles can be generated with dna stored in tris, 0.
Our results showed comparable dna fragmentation index dfi after treatment with both dgc products. Sperm dna fragmentation around 50% of most conditions of infertility could be connected to a male element. Assessment of apoptosis and necrosis by dna fragmentation. Human sperm dna fragmentation and its correlation with. Alternative protocol for dna fragmentation analysis procedure 1. As an alternative, to gel based analysis, consider using a tunel assay kit for the ability to analyze dna fragmentation by flow cytometry or microscopy.
It is known that apparently morphologically normal sperm from men with poor semen quality are at higher risk of being aneuploid10 so the usual approach of choosing sperm for injection using motility and. Dna fragmentation index dfi in at least 30% of male infertility cases, the true cause of infertility remains unknown. Dna is a rich source of information that can be applied to the diagnosis and prognostication of many physiological and pathological conditions such as. Role of sperm dna fragmentation in male factor infertility. Methods to detect chromatin cleavage include tunel assays for whole cells and paraffin sections, dna fragmentation assays using whole cells, assays of total genomic dna, analysis of dna fragmentation by agarose gel electrophoresis, phenol extraction of dna for analysis of fragmentation, a quantitative assay for dna fragmentation, and detection. Sperm contains half of the dna required to fertilize an egg.
Sperm dna fragmentation index and pregnancy outcome after. However, in both cases, a comparable number of samples do not benefit from the treatment. Dna fragmentation index, intracytoplasmic sperm injection, intrauterine insemination, male infertility, sperm function tests, terminal deoxynucleotidyl. Dna damage, such as fragmentation and denaturation, can have adverse effects on fertilization and embryo development and can cause infertility 5. In our prospective study, a total of 1 010 subfertile men, aged from 18 to 50 years old, were enrolled from august 2012 through june 2015. Critical evaluation of two models of flow cytometers for. Sperm dna fragmentation is significantly higher in infertile men and while men with poor semen parameters are more likely to have high sperm dna fragmentation, high sperm dna fragmentation is also found in men with normal semen parameters who may be diagnosed with unexplained infertility 1. Semen analysis forms the basic investigation for male factor of infertility. Esteves, md, phd director, androfert campinas, brazil slideshare uses cookies to improve functionality and performance, and to provide you with relevant advertising.
Apoptosis dna fragmentation analysis protocol abcam. Dna fragmentation index dfi as a measure of sperm quality and fertility in mice. Pdf chapter 2 sperm dna fragmentation researchgate. To summarise the latest evidence on the role of sperm dna fragmentation sdf in male factor infertility, as sdf has been emerging as a valuable tool for male infertility evaluation. Dna fragmentation is a specific marker for apoptosis. Antioxidants to reduce sperm dna fragmentation reproductive. With this assay a dna fragmentation index dfi and high dna stainability hds is calculated and. The place of sperm dna fragmentation testing in current. Therefore, the integrity of sperm dna fragmentation is not only the effective evaluation index to evaluate the quality of sperm, but also the independent clinical and. Each blinded sample, 6 per semen sample, was analyzed using the sperm dna fragmentation assay acridine orangeflow cytometry sdfa and the osa test which directly measures sperm damage from oxidative stress by quantifying the presence of adducts, molecules in semen covalently modified by free radicalsreactive oxygen species. Exalphas dna fragmentation detection kit components are shipped on cold pack. Sperm dna fragmentation and standard semen parameters in.
Sperm dna fragmentation index with unexplained recurrent. Major mechanisms of inducing dna damage in spermatozoa during either the production or the transport of sperm cells. Sperm dna fragmentation index as a promising predictive tool for. Dna fragmentation testing is a type of male fertility test which measures the amount of damaged dna in a sperm sample. Assessment of sperm dna fragmentation sdf has been proposed to. Dna fragmentation detection kit exalpha biologicals inc. Couples with 30% damage are more likely to have success with icsi than ivf. According to who manual sperm values of normalcy are. The present study aimed to assess the relationship between sperm motility, sperm dfi, and rates of clinical pregnancy by analyzing data from 3,000 couples dealing with infertility.
Dna fragmentation is the separation or breaking of dna strands into pieces. Sperm dna fragmentation refers to the presence of sperm with damaged broken dna, which affects male fertility. Sperm dna fragmentation causes, symptoms and treatments. However, it is little known that the correlations of sperm dna damage with obesityassociated markers, and reproductive hormones and lipids levels in serum and seminal plasma. Variations and polymorphisms in genes playing key roles in procedures regulating genome integrity, meiotic recombination, gametogenesis, ie. Numerous techniques for identifying and measuring dna fragmentation have been developed as a means of detecting cell death, including the dna ladder assay, comet assay, and terminal deoxynucleotidyl transferase deoxynucleotide dutp nick end labeling tunel assay. The scsa bases its results on 1 the dna fragmentation index dfi, which is the percent age in the sample that have measurable increased. Analysis of human sperm dna fragmentation index dfi.
Human sperm nuclear dna fragmentation assays and their values in assisted conception mahmood morshedi, ph. The biology of cellfree dna fragmentation and the roles. Methods a search of pubmed was conducted using the keywords sperm dna fragmentation and male infertility. Metaanalysis of sperm dna fragmentation using the sperm. More recent research has suggested that sperm dna fragmentation index dfi may be a more robust and reliable means of predicting assisted reproductive outcomes. This semiquantitative method is a simple technique that provides a robust answer. After fragmentation, dna pieces mix randomly in the dna buffer solution and lose all information regarding spatial organization. It represented the ratio of denatured sperm dna in the total dna. Dna fragmentation, male infertility, sperm aneuploidy, tunel assay. Dna fragmentation sext after undertaking the sperm count, snap freeze and store the raw semen in dry ice or liquid nitrogen within 1 hour of ejaculation in cryovials 0. It thus seems that an increase in dna fragmentation primarily affects in vivo fertility, either by reducing natural conception or by a significant reduction in successful intrauterine inseminations. The dna fragmentation index dfi value was calculated by casawljw9000. Dna fragmentation, though this needs more investigation16.
A semen evaluation that calculates sperm concentration, motility as well as morphology is classically long been utilized like the gold standard evaluation for deciding a mans fertility. Importance of a semen analysis report for determining the. Dna fragmentation index dfi as a measure of sperm quality and. All men have some amount of damage to their sperm dna, but high percentages of damage may indicate greater difficulty achieving pregnancy with iui and ivf. The classical method to detect dna ladders is to examine fragmented genomic dna on an agarose gel. Dna fragmentation is considered a biochemical hallmark of apoptosis. Given that basal levels of reactive oxygen species ros are higher in cancer cells, there is a growing school of thought that endorses prooxidants as potential chemotherapeutic agents.
Terc knockout mice had a six fold mean increase in the percentage of sperm cells with fragmented dna. Predictive value of oxidative stress testing in semen for. Application of digital pcr for assessing dna fragmentation. If the dna in a sperm is damaged, it may be either unable to fertilize the egg, or the resulting embryo will fail to progress and properly implant into the uterine lining. Alterations of any kind in sperm dna are likely to cause infertility in the man affected, as integrity of sperm dna is key to obtaining viable embryos and subsequently a healthy baby. The aim of this study was to investigate whether seminal oxidation. Tips and tricks for optimizing sperm preparation and. While longer, more contiguous reads do preserve more spatial relatedness and thus improve. Total reactive antioxidant potential and dna fragmentation index as fertility sperm parameters. Conventional semen analyses have limitations in male infertility diagnosis and prognosis.
459 1312 174 1459 1561 264 1577 933 745 1126 470 830 1235 1359 1279 1576 781 1033 953 1411 194 1422 1013 429 941 1509 166 572 560 568 763 873 550 1094 520 1097 416