Chronic myelogenous leukemia (CML) is a myeloproliferative neoplasm arises from Bcr-Abl gene translocation(called Ph chromosome) in hematopoietic stem cells (HSCs).JAK2V617F mutation is an acquired singlenucleotide polymorphism (SNP) occurs in JAK2 gene and is associated with many hematological malignancyother than CML. This study aimed to investigate the prevalence of JAK2V617F mutation and serum levels ofalkaline phophatase (ALP) and lactate dehydrogenase (LDH) in Ph+ CML Iraqi patients treated with imatinib.Blood samples were collected from 42 Ph+ CML patients who have been received at least six month therapywith imatinib. DNA was extracted, and real time polymerase chain reaction (qPCR) was used for JAK2V617Fdetection. Serum levels of ALP and LDH were measured using ready kits. Five of 43 CML patients (11.62%)had heterozygous mutant allele of JAK2V617F mutation, with a concentration ranged from 0.01% to 0.12%. Theprevalence of this mutation is more associated with male than female (OR=0.5, 95%CI=0.364-0.687).JAK2V617F-positive patients had higher average serum levels of ALP and LDH (146.05±8.028 IU/L and 204±10.85 IU/L respectively) than that of JAK2V617F-negative patients (64.45±40.15 IU/L and 178.33±13.693 IU/Lrespectively with significant differences. JAK2V617F mutationcould occur in coexistence with Bcr-Abl transcriptin CML patients, and serum levels of ALP and LDH can be used as indicators for this coexistence .