Studies have explored the influence of DNA damage in assisted reproductive technology (ART), but the outcome remains controversial. To determine whether sperm DNA fragmentation index (DFI) has any effect on ART outcomes, we collected detailed data regarding 1,333 IVF cycles performed at our centre, and the data of our retrospective cohort study were extracted for this meta‐analysis. We searched PubMed, Web of Science, EMBASE and Google Scholar and performed a systemic review and meta‐analysis. Primary meta‐analysis of 10 studies comprising 1,785 couples showed that live birth rate was no significantly different between low‐DFI group and high‐DFI group (p > 0.05). Secondary meta‐analysis of 25 studies comprising 3,992 couples showed a higher miscarriage rate in high‐DFI group than in low‐DFI group (RR=1.57 [1.18, 2.09], p < 0.01). Meta‐analysis of eight studies comprising 17,879 embryos revealed a lower good‐quality embryo rate (RR=0.65 [0.62, 0.68], p < 0.01). Meta‐analysis of 23 studies comprising 6,771 cycles showed that the high‐DFI group had a lower clinical pregnancy rate than low‐DFI group (RR=0.85 [0.75, 0.96], p < 0.01). Heterogeneity of included studies weakened our conclusions. Our study showed that DFI has adverse effects on ART outcome. More well‐designed studies exploring the association between DFI and ART outcome are desired.