Analytica chimica acta · 2025
Precise detection of sperm DNA breaks and AP sites based on TDT and strand displacement probes.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод в очереди
Обработка статьи · этапы, ошибки и управление
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Yan Bei, Liu Ziyang, Fan Heng, Wang Juan, Zhang Fan, Pei Liguo, Liu Xiaowen, Ma Lianghong, Xiao Xianjin
Аннотация
BACKGROUND: Sperm DNA damage is an important factor associated with male infertility, miscarriage, and adverse health outcomes in offspring. Current detection methods face challenges such as inaccurate assessments, limited evaluation indicators, and poor reproducibility, largely due to design flaws and constraints in measurable parameters. Therefore, there is an urgent need to develop more reliable detection technologies and to establish more precise and comprehensive evaluation standards to address this critical issue.
RESULTS: This study developed an innovative biosensor based on terminal deoxynucleotidyl transferase (TdT) and strand displacement (SD) probes, establishing a method for the simultaneous detection of two key biomarkers of sperm DNA damage: DNA breakpoints and apurinic/apyrimidinic (AP) sites. By incorporating standardized reference controls, the assay significantly improved detection accuracy and reproducibility, thereby enhancing clinical assessment efficacy. The method was applied to analyze sperm samples from 40 men with normal semen parameters, 50 asthenozoospermia patients, 48 fertile individuals, and 44 subjects with recurrent pregnancy loss (RPL). Receiver operating characteristic (ROC) curve analysis demonstrated that the combined detection of the mean number of sperm DNA breakpoints (MDB) and AP sites significantly outperformed individual assessments of MDB, DNA fragmentation index (DFI), or AP alone in predicting RPL, exhibiting superior sensitivity and specificity.
SIGNIFICANCE: This advanced technology enables a more comprehensive and precise quantification of sperm DNA damage, overcoming the limitations of traditional methods that have singular assessment indicators, poor effectiveness, and low reproducibility. In addition to demonstrating better clinical relevance in the diagnosis of recurrent pregnancy loss (RPL) and asthenozoospermia, this approach also provides a powerful tool for research in male infertility and holds significant promise for clinical applications.
RESULTS: This study developed an innovative biosensor based on terminal deoxynucleotidyl transferase (TdT) and strand displacement (SD) probes, establishing a method for the simultaneous detection of two key biomarkers of sperm DNA damage: DNA breakpoints and apurinic/apyrimidinic (AP) sites. By incorporating standardized reference controls, the assay significantly improved detection accuracy and reproducibility, thereby enhancing clinical assessment efficacy. The method was applied to analyze sperm samples from 40 men with normal semen parameters, 50 asthenozoospermia patients, 48 fertile individuals, and 44 subjects with recurrent pregnancy loss (RPL). Receiver operating characteristic (ROC) curve analysis demonstrated that the combined detection of the mean number of sperm DNA breakpoints (MDB) and AP sites significantly outperformed individual assessments of MDB, DNA fragmentation index (DFI), or AP alone in predicting RPL, exhibiting superior sensitivity and specificity.
SIGNIFICANCE: This advanced technology enables a more comprehensive and precise quantification of sperm DNA damage, overcoming the limitations of traditional methods that have singular assessment indicators, poor effectiveness, and low reproducibility. In addition to demonstrating better clinical relevance in the diagnosis of recurrent pregnancy loss (RPL) and asthenozoospermia, this approach also provides a powerful tool for research in male infertility and holds significant promise for clinical applications.