Introduction: To explore the prevalence of antisperm antibodies (ASA) and its impact on the embryo culture outcomes of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI), a retrospective analysis of the Center for Reproductive Medicine’s data was conducted. Methods: 8,265 semen samples were detected for ASA using mixed antiglobulin reaction. In addition, 934 IVF/ICSI cycles were included to compare the effects of ASA on embryo culture outcomes during IVF/ICSI, including normal fertilization rate, D3 good-quality embryo rate, available blastocyst formation rate, and available embryo rate. Results: The positive rate of ASA in semen was relatively low, about 1.03%. We found ASA may weaken the normal fertilization rate of IVF cycles. In addition, there was no statistical difference in outcomes between the positive groups and negative groups of ICSI cycles with a threshold 10% in ASA, but ASA significantly reduced the normal fertilization rate and D3 good-quality embryo rate of ICSI cycles with a threshold 50% in ASA. It was speculated that the high ratio of rescue ICSI cycles resulted in the poor embryo culture outcomes. We supposed that the higher of the ASA-positive rate, the higher of the likelihood of fertilization failure of IVF. Comparing with IVF, ICSI is more recommended as a fertilization method for ASA-positive patients who need assisted reproductive technology. Conclusions: ASA may have a negative impact on the embryo culture outcomes of IVF. Therefore, for ASA-positive patients who require IVF/ICSI treatment, it is suggested embryologists wash the sperm thoroughly and use short-term fertilization of IVF or conventional ICSI to assist with pregnancy.

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