Whether the prophylactic use of antibiotics increase the risk of necrotizing enterocolitis (NEC) remains controversial. This review aims to investigate initial empirical antibiotic therapy (IEAT) and is associated with the risk of NEC. PubMed, EMBASE, Cochrane Library, and Web of Science databases were searched through March 1, 2020. All studies on the impacts of antibiotic exposure on NEC development were included. Thirteen studies including 7901 participants were selected. Two reviewers independently examined the extracted data and assessed the quality of the included studies. Random-effects model was used to pool the effect estimates. We found that IEAT (≥ 5 days) was associated with an increased risk of NEC in adjusted (Odds risk [OR] 1.51, 95% confidence interval [CI] 1.22-1.87) and unadjusted (OR 2.35, 95% CI 1.54-3.57) analyses. Sensitivity analysis also supported these findings.Conclusion: The evidence suggests an association between IEAT (≥ 5 days) and the risk of NEC. Further studies are needed to address whether the association with IEAT is causal.What is Known:•Necrotizing enterocolitis (NEC) is acute inflammatory necrosis of the intestinal tractin the newborn infant.•Some observational studies have associated initial empirical antibiotics with an increased risk of subsequent NEC.What is New:•Initial empirical antibiotic therapy (IEAT) (≥ 5 days) appear to increase the risk of NEC.