Neonatal septicemia (NS) is a common cause of death of newborn infants, hence early diagnosis and treatment are of the utmost importance. However, lack of specific clinical symptoms and late detection delay a correct diagnosis. It is therefore of great importance to establish auxiliary indexes for the early diagnosis of NS. To evaluate the value of interleukin (IL-6 and IL-8) in the diagnosis of NS, a prospective study was carried out. Seventy-five newborns who developed septicemia and received treatment in our hospital from January 2013 to December 2014 were selected as research subjects; also, 50 healthy newborns were set as a control group. The levels of serum IL-6 and IL-8 were compared between the two groups. Results demonstrated that levels of C-reactive protein (CRP), IL-6 and IL-8 of the septicemia group were higher than those of the control group on admission, although the difference had no statistical significance (P less than 0.05); the septicemia group had higher sequential organ failure assessment (SOFA) scores but lower pediatric critical illness scores (PCIS) compared to the control group (P less than 0.05); levels of CRP, IL-6 and IL-8 were in positive correlation to the SOFA scores and in negative correlation to PCIS. Analysis of receiver operating characteristics (ROC) curve demonstrated that the sensitivity, specificity and accuracy were 85.7%, 80.2% and 81.8%, respectively, when IL-6 level was set as 32 pg/mL, 78.1%, 64.2% and 66.9%, respectively when IL-8 level was set as 54 pg/mL, and 71.4%, 86.3% and 82.7% respectively, when detection of IL-6 and IL-8 were combined together. Hence it can be concluded that: IL-6 and IL-8 are involved in inflammatory reactions; levels of IL-6 and IL-8 were correlated to the severity of the infection; the value of IL-6 is higher than that of IL-8 in the diagnosis of neonatal septicemia and the combined detection of IL-6 and IL-8 can improve the accuracy of the diagnosis of neonatal septicemia.