Background Bloodstream infection (BSI) is a life-threatening condition caused by the presence of microorganisms, generally caused by a range of bacteria in the blood. Objectives The aim of this study was to evaluate the possible role of procalcitonin (PCT) and C-reactive protein (CRP) as biomarkers of pediatric BSI. Methodology The study was conducted on 150 blood samples collected from the patient who admitted to Children Welfare Teaching Hospital, Medical City, Baghdad. During the period from November 2020 to March 2021, ninety blood samples from them were positive culture and 60 blood samples were negative culture (control group). The isolates were identified depending on the morphological, microscopic examination, and biochemical tests. Moreover, serum was obtained from all participants for the determination of the screening level of human PCT measured by enzyme-linked immunosorbent assay and CRP by slide agglutination test. Results The results in this study revealed that the mean levels of serum PCT and CRP in Gram-negative group and Gram-positive group were significantly difference from the control group. Conclusions The adoption of these biomarkers as routine diagnostic tests for sepsis may help in the early diagnosis of pediatric sepsis.