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Abstract Introduction: Neonatal sepsis represents a major cause of mortality in developing countries.The gold standard for establishing a diagnosis is through culture. However, several factors can make the confirmation of sepsis a diagnostic challenge.This study aimed to measure serum interleukin-27 (IL-27) and mean platelet volume (MPV) in neonates diagnosed with late-onset sepsis (LOS). Subjects and Methods: This cross- sectional study was conducted on 90 full term newborns admitted to the Neonatal Intensive Care Unit of Cairo University Hospitals from June 2019 to January 2020. Neonates were assigned into two equal matched groups: LOS group and control group. All neonates were subject to full history taking and detailed clinical assessment. Laboratory data including complete blood picture with MPV and blood culture results were documented as well as highly sensitive C-reactive protein (CRP) levels. Serum IL-27 levels were measured using enzyme-linked immunosorbent assay. Results: Clinical findings suggestive of sepsis and most complete blood picture parameters as well as highly sensitive CRP showed significant differences between the two groups (P < 0.05). The median level of IL-27 and mean level of MPV were significantly higher in LOS group as compared to the control group (4364 vs. 4.5 pg/ml, P < 0.001 and 12.02 ±1.54 FL vs. 9.42 ± 1.08 FL, P < 0.001, respectively). At a cut-off value of 283.8 pg/ml, IL-27 showed a diagnostic sensitivity of 97.8% and a specificity of 100%. While at a cut-off value of 11.55 FL, the MPV had a sensitivity of 77.8% and a specificity of 97.8% |