Objective: To investigate the relationship between single nucleotide polymorphisms of pulmonary surfactant protein B gene and respiratory distress syndrome in premature infants. Methods: Ninety-two preterm infants with neonatal respiratory distress syndrome (NRDS) were selected as the NRDS group. Another 92 cases without respiratory distress syndrome were selected as the non-NRDS group. The single nucleotide polymorphisms of SPB-18 A/C locus and SPB-1580 C/T locus were detected in the groups, and the genotype distributions of the SPB-18 A/C locus and the SPB-1580 C/T locus were compared between the two groups. The relative risk of both sites and the onset of respiratory distress syndrome were analyzed using an chi(2) test. Results: In the SPB-18 A/C locus genotype test, AA, CC, and AC in the NRDS group were 20.65%, 35.87%, and 43.47%, respectively, which were not significantly different from 19.57%, 32.61%, and 47.83% in the non-NRDS group (P = 0.854, P = 0.641 and P = 0.554). In the SPB-1580 C/T locus genotype test, CC in the NRDS group accounted for 66.30%, which was higher than the corresponding value in the non-NRDS group (40.22%, P = 0.000). T in the NRDS group accounted for 5.43%, and CT accounted for 28.26%, which were lower than the corresponding values in the non-NRDS group (14.13%, P = 0.047; 45.65%, P = 0.015). SPB-18 A/C was not associated with NRDS in preterm infants (P = 0.833), whereas the SPB-1580 C/T site was associated with respiratory distress syndrome in preterm infants (P = 0.000). Respiratory distress syndrome in preterm delivery of C allele with SPB-1580 locus was 2.410 times higher than it was in preterm infants with a confidence interval of (1.504, 3.862). Conclusion: The SPB-1580 A/C polymorphism is associated with respiratory distress syndrome in preterm infants. Carrying the C allele of SPB-1580 may be a risk factor for respiratory distress syndrome in preterm infants.