Impact of Post Wash Incubation Time on Sperm DNA Integrity in Normal and Abnormal Samples

Sperm DNA fragmentation is increasingly recognised as a critical factor affecting male fertility, especially in Assisted Reproductive Technology ART like intrauterine insemination IUI. While sperm preparation methods such as swim-up and density gradient centrifugation help reduce initial DNA damage, the removal of seminal plasma may render spermatozoa more vulnerable to oxidative stress over time and this raises concern about stability of DNA integrity during common delays between sperm preparation and insemination. Existing studies have assessed sperm DNA fragmentation at a single time point or over short intervals, with limited data available on its progression beyond 3 hours post-processing. Moreover, these studies lack standardization and do not differentiate between normal and abnormal semen samples which is referenced by WHO 2021 guidelines. This study aims to evaluate DNA fragmentation at 0 hour, 4 hour and 24 hour after sperm preparation using sperm chromatin dispersion test SCD assay in samples to determine timing for clinic use. The assay will be carried out in department lab with in house preparation.