The current study aimed at detection of leukemia stem cells (LSCs) expressing CD19, CD34 and CD38 (CD19+/CD34+/CD38Low) by triple color flow cytometry before and after induction chemotherapy in 20 newly diagnosed pediatric B-ALL patients. LSCs were detected in 16/20 (80%) of cases at presentation. The mean value of LSCs was 1.57 ± 4.16% at presentation and 0.18 ± 0.34% after induction therapy. No statistically significant difference was encountered between the percentage of LSCs before and after induction chemotherapy, denoting that these cells resist the traditional lines of treatment used for childhood B-ALL. Although LSCs were encountered in 14/20 (70%) of patients after induction chemotherapy, all patients achieved complete remission after induction, while two died from progressive disease in the follow up period for one year. So, we recommend the estimation of LSCs expressing CD19, CD34 and CD38Low in de novo cases of ALL before starting treatment and in the follow up period to monitor the response to therapy, for detection of minimal residual disease and early detection of relapse and in the future, it may be a target for therapy.