Research question: Is the pregnancy rate affected by artificial oocyte activation (AOA) with A23187 after intracytoplasmic sperm injection (ICSI) in infertile patients?Design: Our retrospective study included 308 patients who transferred blastocyst from routine intracytoplasmic sperm injection (ICSI) and 82 patients who transferred blastocyst from routine ICSI combined with AOA (ICSI-AOA) from January 2014 to April 2020. Pregnancy outcomes of couples who received routine ICSI or ICSI-AOA were analyzed after the first blastocyst transfer, which covered frozen-thawed blastocyst transfer and fresh blastocyst transfer. AOA was performed with A237817. We used multivariable logistic regression analysis to determine which variables could be independently associated with the pregnancy rate. Effect sizes were summarized as odds ratios (ORs), with precision evaluated by 95% CIs. Results: The clinical pregnancy rate was 71.95% in the AOA group and 57.47% in the routine ICSI group. The effect size of the AOA on clinical rate was evaluated in prespecified and exploratory subgroups in each subgroup. And multivariable logistic regression analysis was performed to identify factors associated with the clinical rate. The AOA group had a higher chance of clinical pregnancy in all subgroups: female age at oocyte retrieval, female BMI, protocol in the fresh cycle, female infertility type, MII oocyte numbers, fresh or frozen blastocyst transfer, No. of blastocyst transfer and blastocyst quality. Multivariable analysis showed AOA to be associated with an increased likelihood of clinical pregnancy compared with routine ICSI (p=0.03; adjusted OR 1.89, 95% CI 1.09–3.27).Conclusions: This study suggested that AOA can increase the rate of clinical pregnancy obviously, which helps clinicians to advise patients on AOA risks.