Background: Neonatal necrotizing enterocolitis (NEC) is a serious pediatric gastrointestinal disease and a cause of death in neonates, especially in premature infants. The addition of probiotics to the diet can reduce the incidence and severity of neonatal NEC. This meta-analysis explored the preventive effect of probiotics on NEC. Methods: Endnote X9 software was used to search for relevant studies in the Ovid, Embase, PubMed, and Web of Science databases. The search terms were "probiotics" and "necrotizing enterocolitis". After retrieval, screening, and quality evaluation of the studies, Stata 16.0 software was used to analyze the data. Results: A total of 10 studies, which collectively included 3,227 patients, were selected for analysis. Of them, 5 used a multiple-strain probiotics, and 5 used single-strain probiotic. Meta-analysis showed that treatment with probiotics could reduce the incidence of severe NEC [risk ratio (RR) =0.66; 95% confidence interval (CI): (0.50, 0.87); Z=-2.978; P=0.003], reduce mortality in underweight premature children [RR =0.81; 95% CI: (0.70, 0.94); Z=-2.864; P=0.004], and reduce the incidence of feeding intolerance [RR =0.78; 95% CI: (0.67, 0.90); Z=-3.280; P=0.001]. Discussion: The addition of probiotics to the diet of low-birthweight and premature infants can reduce the incidence of severe NEC and reduce related mortality rates.