The CCAAT/enhancer-binding protein-alpha (CEBPA) is lineage-specific transcription factor in the hematopoietic system. Several studies have reported the presence and frequency of CEBPA mutations.In this study, we aimed the clinical features and the prognostic significance associated with CEBPA mutations in 30 pediatric patients with acute leukemia.In addition, association between found variants and mutations of Ten-Eleven-Translocation 2 (TET2), Kirsten rat sarcoma viral oncogene homolog (KRAS), and Casitas B-cell lymphoma (CBL), Fms-Related Tyrosine Kinase (FLT3), Januse Kinase-2 (JAK2) and Nucleophosmin 1 (NPM1) were analyzed which are important prognostic risk factors for pediatric acute leukemia. The entire CEBPA coding region was screened using the Next Generation Sequencing (NGS) method. Furthermore, TET2, KRAS and CBL genes were screened using the NGS method, FLT3 gene was analyzed by Real Time Polymerase chain Reaction (Real Time-PCR).JAK2 and NPM1 genes were screened by Sanger DNA sequencing. CEBPA mutations were detected in 16 (53.3%) of 30 patients. In total, ten distinct of nucleotide changes were identified in 30 patients including 6 novel and 4 known mutations by sequencing the entire CEBPA gene. We found 6 frame shift mutations, 1 missense mutation, 3 synonymous variant. The most common mutation was the c.487del G resulting p.Glu163Ser in 5 cases. Three patients carried CEBPA double mutations. The detected variants in this article seem to be the first screening results of genes studied by NGS in pediatric acute leukemia patients. Our results also showed some degree of association between FLT3-ITD, TET2, KRAS, CBL and CEBPA mutations.