Objective To explore the risk factors of spontaneous intestinal perforation (SIP) in premature infants. Methods A retrospective case-control study was designed. Preterm infants diagnosed of SIP and admitted from January 2010 to June 2020 were included in the SIP group. Preterm infants of the same gestational age and without intestinal perforation during the same period were randomly selected as the control group. The clinical characteristics between the two groups were analyzed and compared, and the risk factors of SIP were analyzed. Results A total of 21263 preterm infants were admitted during the study period, including 28 infants ( 14 boys and 14 girls) in the SIP group. The perforation age was 8 . 5 ( 2 . 5 - 11 . 0 ) days, gestational age was ( 32 . 9 ±2 . 5 ) weeks, and the birth weight was ( 1831 . 5 ±475 . 2 ) g. At the same time, 61 infants ( 36 boys and 25 girls) were included in the control group, with gestational age of ( 33 . 2 ±2 . 6 ) weeks and birth weight of ( 1869 . 5 ±493 . 8 ) g. Univariate analysis showed that there were statistically significant differences between the SIP group and the control group in the proportion of periventricular-intraventricular hemorrhage (PIVH) of grade Ⅲ and above, early-onset sepsis, septic shock, hypoproteinemia, anemia and dopamine exposure within one week (all P< 0 . 05 ). Conclusion PIVH grade Ⅲ and above, earlyonset sepsis, septic shock, hypoproteinemia, anemia, and dopamine exposure within one week may be related to SIP.