AIM: To understand the efficacy and safety of amikacin (AMK) for the treatment of carbapenem-resistant Klebsiella pneumoniae pneumoniae (CRKP) in preterm infants and to establish a management process for the use of amikacin in preterm infants. METHODS: CRKP-infected preterm infants treated with amikacin between January 2019 and December 2021 were retrospectively analyzed, and parametric data paired t-tests were used to assess the efficacy and safety of amikacin for the included infectious and safety indicators, and to establish a management process for amikacin use in preterm infants. RESULTS: Eight cases of CRKP infection were included, with the main diagnosis of pneumonia and sepsis. eight preterm infants were screened for the AMK ototoxicity gene mitochondrial gene MT-RNR1 (MT-RNR1 1494C>T and MT-RNR11555A>G) before amikacin treatment, and none of them were found to have the gene variant. after receiving amikacin sulphate injection treatment for 7 days, the indicators of infectivity were improved, and was statistically significant (P<0.01). No clinical ototoxicity or nephrotoxicity was observed in the children before or after treatment. CONCLUSION: Aminoglycosides are still the main antibiotics used for the empirical treatment of suspected infections in preterm infants, especially drug-resistant bacterial infections. Despite the risk of ototoxicity and nephrotoxicity, we provide management procedures and recommendations for neonatal treatment with amikacin to reduce the risk of ototoxicity and nephrotoxicity in AMK.