Chronic myelogenous leukemia (CML) is a myeloproliferative neoplasm arises from Bcr-Abl gene translocation (called Ph chromosome) in hematopoietic stem cells (HSCs). This genetic abnormality results in constitutive activation of tyrosine kinase and subsequent uncontrol growth and multiplication of granulocytes. The cornerstone in treatment of CML are tyrosine kinase inhibitors, of which imatinib is the most effectively used. JAK2V617F mutation is an acquired single nucleotide polymorphism (SNP) occurs in JAK2 gene and is associated with many hematological malignancy other than CML. It was thought that the two genetic abnormalities (Bcr-Abl and JAK2V617F) occur mutually; however, growing body of evidences suggested the reverse. This study aimed to investigate the prevalence of JAK2V617 mutation associated with serum levels of alkaline phophatase (ALK) and lactate dehydrogenase (LDH) in Ph+ CML Iraqi patients treated with imatinib. A total of 43 Ph+ CML patients (24 males and 18 females, age range 16-80 years) who attend Iraqi National Center of Hematology for Research and Treatment/Baghdad were enrolled in this study. Each patient has been received at least six month therapy with imatinib. A consent form involving age, gender, height, weight, smoking status, residency and first family relative history of leukemia was obtained from each patient. Besides, blood samples were collected, from which the granulocytes were separated and then DNA was extracted using a ready kit. Two assays were used for detection of JAK2V617F mutation; real time polymerase chain reaction (qPCR) using specific primers and probe, and allele specific PCR (AS-PCR) using specific primers. Total white blood corpuscles (WBC) as well as serum levels of ALP and LDH were measured. qPCR assay revealed 5 patients out of 43 (11.62%) were heterozygous for the muatant allele of JAK2V617F mutation (genotype GT). The concentration of this allele ranged from 0.01% to 0.12%. None of blood sample gave positive result for AS-PCR assay. From the all risk factors, only gender had significant association with the incidence of JAK2V617F mutation (p= 0.034, OR= 0.5, 95%CI= 0.364-0.687). Average total WBC count, and serum levels of ALP and LDH were higher in JAK2V617F-positive patients (9042±1512.55, 146.05±8.028 IU/L and 204±10.85 IU/L respectively) than that of JAK2V617F-negative patients (6039±1772.239, 64.45±40.15 IU/L and 178.33±13.693 IU/L respectively) with significant differences. These results indicate that JAK2V617F mutation can occur simultaneously with Ph chromosome in CML patients, and qPCR is a highly sensitive method for the detection of this mutation. Furthermore, serum activity of APL can be used as an indicator for the presence of JAK2V617F mutation in CML patients.