Abstract: Objective　To analyze the risk factors of neonatal respiratory distress syndrome (NRDS) in twins. Methods Medical record of twins charged in Children's Hospital of Chongqing Medical University between 2005 and 2016 were retrospectively reviewed. Twins with one of sibling suffered from NRDS were enrolled and were divided into NRDS group and non-NRDS group, and infants were divided into less than 34 weeks group and more than 34 weeks group according to their gestational age. The potential risk factors of NRDS including gender, birth weight, delivery order, amniotic fluid contamination, premature rupture of membranes, Apgar score at 1 and 5 minutes after birth were compared between two groups. Results　A total 72 pairs of twins were recruited and no difference of gender and birth weight were found between NRDS and non-NRDS groups (P >0.05). Rate of second-born twins (66.7% vs. 33.3, χ2=16, P =0.000) and lower Apgar score at 1 minute after birth (41.7% vs.22.2%, χ2=6.261, P =0.012) were significantly higher in twins with NRDS than those without NRDS. However, there was no significant difference of Apgar score at 5 minute between NRDS and non-NRDS groups (P>0.05). For twins with more than 34 weeks of gestational age, there were no statistically significant difference of birth order and the Apgar score at 1 minute between NRDS and non-NRDS groups (P>0.05). For twins less than 34 weeks of gestational age, the rate of second-born and lower Apgar score at 1 minute in NRDS group were higher than that in non-NRDS group (P<0.05). Logistic regression analysis showed that second-born twins was associated with higher incidence of NRDS (OR=4, 95%CI: 2.00~8.00). Conclusions　The second-born twins with less than 34 gestational age had increased odds of RDS.