article · Current Issues in Molecular Biology
Assessing male fertility and managing assisted reproductive technology requires effective, non-invasive indicators of reproductive health. Seminal cell-free DNA, easily quantified in biological fluids, offers insight into cellular apoptosis and necrosis within the male reproductive tract. Measuring these DNA fragments via quantitative PCR targeting the RNase P gene revealed distinct concentration differences across various severe sperm abnormalities compared to normozoospermic controls. Seminal plasma concentrations were significantly higher in men with total azoospermia and very severe teratozoospermia than in healthy controls, even after statistical adjustments for multiple testing. Other conditions, such as very severe asthenozoospermia and oligozoospermia, did not retain statistically significant elevations. These findings indicate that seminal cell-free DNA levels rise markedly in the most pronounced forms of spermatogenic impairment, highlighting the biomarker potential of cell-free DNA for non-invasive evaluations of sperm quality.
Evaluating male infertility often relies on conventional semen analysis, which may not capture underlying testicular damage or cell death. Demonstrating that cell-free DNA in seminal fluid correlates specifically with severe conditions such as azoospermia provides a potential non-invasive diagnostic tool. This biological marker could help clinicians better identify cellular impairment in patients undergoing assisted reproduction.
This research is at an early stage and could eventually support non-invasive diagnostic assays for fertility clinics and assisted reproductive technology laboratories. Development into a clinical test will require standardising reference thresholds and conducting larger multicentre, longitudinal trials to confirm diagnostic accuracy across wider patient populations.
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Cell-free DNA (cfDNA) fragments represent emerging biomarkers of major interest in the assessment of male fertility and the management of couples undergoing assisted reproductive technology (ART). Easy to quantify and present in most biological fluids, seminal cfDNA could reflect apoptosis and necrosis occurring within the male reproductive system. In this study, we compared cfDNA concentrations, measured by means of qPCR targeting the RNase P gene, in the seminal plasma of men with severe sperm quality abnormalities, including very severe asthenozoospermia (n = 25), total azoospermia (n = 25), very severe oligozoospermia (n = 25), and very severe teratozoospermia (n = 25), with those of normozoospermic controls (n = 25). Mean cfDNA concentrations were particularly elevated in men with azoospermia (5.47 ± 1.17 µg/mL) and very severe teratozoospermia (3.25 ± 1.21 µg/mL), compared with controls (1.96 ± 0.27 µg/mL). After adjustment for multiple testing, only azoospermia and very severe teratozoospermia remained significantly associated with increased cfDNA levels (Holm–Bonferroni adjusted p < 0.003), whereas the other phenotypes did not maintain statistical significance. These results suggest that seminal cfDNA is markedly increased in the most severe forms of spermatogenic impairment, potentially reflecting enhanced germ cell apoptosis or testicular cell involvement. Despite some limitations, including the modest sample size and single-center design, our observations support the value of cfDNA as a promising non-invasive biomarker of sperm quality. Larger multicenter and longitudinal studies are needed to validate its clinical relevance and determine standardized reference thresholds for clinical application.
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DOI: 10.3390/cimb48090911
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