Background Fever is a common presenting complaint to the emergency department (ED), and the evaluation of the febrile child remains a challenging task. Objective The aim of this study was to examine the relationship between secretory phospholipase A2 (sPLA2) and infection in febrile children. Methods A prospective convenience sample of children presenting with fever to an urban pediatric ED were studied. Blood and urine cultures, a complete blood count, and serum concentrations of sPLA2 were obtained, and patients were compared based on their final diagnosis of either a viral or bacterial infection. Results In the 76 patients enrolled, 60 were diagnosed with a viral infection, 14 with a bacterial infection, 1 with Kawasaki disease, and 1 with acute lymphoblastic leukemia. The difference in the serum concentration of sPLA2 in patients with viral infections (22 ± 34 ng/mL) versus those with bacterial infections (190 ± 179 ng/mL) was statistically significant (P < .0001). Receiver operator characteristic curve analysis revealed that sPLA2 was more accurate at predicting bacterial infection (area under the curve = 0.89) than the total white blood cell count (area under the curve = 0.71) and that a value of more than 20 ng/mL had a sensitivity of 93%, specificity of 67%, positive predictive value of 39%, and negative predictive value of 97%. Conclusion Secretory phospholipase A2 differs significantly in children with viral versus bacterial infection and seems to be a reliable screening test for bacterial infection in febrile children.