Purpose: This study aimed to identify the effects of a direct breastfeeding program for premature infants in neonatal intensive care units (NICUs). Methods: This quasi-experimental study was conducted during August 2016 to April 2017. Sixty mothers of premature infants were assigned to the experimental (n = 31) or control groups (n = 29). The program was comprised of breastfeeding education and direct breastfeeding support. The experimental and control groups were provided with education and counseling on breastfeeding at the time of admission and discharge. In the experimental group, the mothers initiated oral feeding with direct breastfeeding and engaged in breastfeeding at least seven times during the NICU stay. The collected data were analyzed by the χ2 -test and repeated measures ANOVA using an SPSS program. Results: The experimental group showed a higher direct breastfeeding practice rate (χ2 = 19.29, p < .001), breastfeeding continuation rate (χ2 = 3.76, p < .001), and self-efficacy (F = 25.37, p < .001) than the control group except for maternal attachment. Conclusion The direct breastfeeding program in the NICU has significant effects on the practice and continuation rate of breastfeeding and breastfeeding self-efficacy. Therefore, this program can be applied in the NICU settings where direct breastfeeding is limited.