期刊
ANALYTICA CHIMICA ACTA
卷 1280, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.aca.2023.341821
关键词
Terminal transferase; Strand displacement probe; Sperm DNA integrity; DNA fragmentation index; Mean number of DNA breaks
A detection system based on terminal deoxynucleotidyl transferase and DNA strand displacement fluorescent probe was developed to quantitatively detect breakpoints at the DNA molecular level. The system showed reasonable correlation and better prediction efficiency compared to the widely accepted SCSA method, and its applicability was proven in assisted reproductive technology procedures.
DNA integrity is crucial for the clinical pregnancy outcome and offspring health, while detection methods currently used (comet assay, TUNNEL assay, SCSA, etc.) can only provide the ratio of positive sperms at the cellular level and are unable to quantitatively detect the breakpoints at the DNA molecular level. Herein, we developed a detection system based on terminal deoxynucleotidyl transferase and DNA strand displacement fluorescent probe, which could efficiently and conveniently measure the number of 3 '-OH (equivalent to the number of breakpoints). We further investigated the use of this technique in assisted reproduction after completing the principle verification, system optimization, and research on analytical performance. The detection system was shown to have a good linear range from 0.01 nM to 4 nM, using single-stranded DNA with 3 '-OH end as the calibrator. The system underwent thorough optimization for stability and accuracy. In comparison to the widely accepted index DFI detected by SCSA, the new system demonstrated reasonable correlation and better prediction efficiency. Its applicability was also proven through its use in assisted reproductive technology procedures.
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