Objective: It is thought that twin neonates born from pregnancies resulting from assisted reproductive technology (ART) are clinically riskier than twin neonates born from spontaneous pregnancy. However, information on the risks in premature infants born as a result of ART pregnancies is limited. In our study, premature twin infants born from ART and spontaneous pregnancies were compared to clinical outcomes. Method: All premature twin infants hospitalized in our unit between September 2017 and September 2019 and born under 32 weeks of gestation were included in our study. Demographic and clinical results of premature twins born as a result of spontaneous and ART pregnancies were compared. Results: A total of 142 premature twins, 116 (81.6%) in the spontaneous twin group and 26 (18.4%) in the ART twin group, were included in the study. Demographic and clinical features were similar between ART and spontaneous twin groups (P > 0.05). Conclusion: Our study determined that premature infants born due to ART pregnancies did not have any additional risk compared to spontaneous infants. This result shows that the main determinants of clinical outcomes in premature infants are gestational week and birth weight.