Objective To compare the application effect of nasal continuous positive airway pressure (NCPAP), nasal intermittent positive pressure ventilation (NIPPV) and high-flow nasal cannula oxygen therapy (HFNC) in the initial treatment of premature infants with respiratory distress syndrome (RDS). Methods A total of 198 premature infants with RDS were randomly divided into NCPAP group (n=65), NIPPV group (n=61) and HFNC group (n=72), and three non-invasive ventilation modes (NCPAP, NIPPV and HFNC) were given respectively. The parameters of blood gas analysis such as pH value, arterial partial pressure of carbon dioxide (PaCO2), respiratory index (RI) and partial pressure of oxygen in arterial blood/fractional concentration of inspiratory oxygen (P/F) were compared among the groups at 24 hours after non-invasive ventilation. The fraction of inspiratory oxygen (FiO2), duration of non-invasive ventilation, adverse reactions and long-term complications were compared among the groups, and the related factors of non-invasive ventilation failure were researched by multivariate Logistic regression analysis. Results Before treatment, there was no statistically significant difference in pH value and PaCO2 value in blood gas analysis among the three groups (all P>0.05). After treatment, there was no statistically significant difference in pH value in blood gas analysis among the three groups (P>0.05), and the PaCO2 level of blood gas analysis in the HFNC group was significantly higher than that in the NCPAP group and NIPPV group (all P<0.05). The RI value and 24 h FiO2 value in the HFNC group were significantly higher than those in the NCPAP group and NIPPV group, while the P/F value was lower than that in the NCPAP group and NIPPV group (all P<0.05). Compared with NCPAP group, the P/F value was higher and the 24 h FiO2 value was lower in the NIPPV group (all P<0.05). There were no statistically significant differences in the duration of non-invasive ventilation, the rate of conversion from non-invasive ventilation failure to invasive ventilation, and the incidence of related complications among the three groups (all P>0.05). The results of multivariate Logistic regression analysis showed that non-invasive ventilation mode and RDS grade were independent influencing factors for the failure of non-invasive ventilation. Conclusion NCPAP or NIPPV is superior to HFNC in improving oxygenation and reducing CO2 retention in premature infants with RDS. The choice of ventilation mode and the severity of RDS are independent influencing factors for the failure of non-invasive ventilation.