EXPLORING JAK2 GENE MUTATION (V617F) THROUGH REAL TIME PCR IN POLYCYTHEMIA VERA PATIENTS IN KPK.

JPUMHS;2024:14:01,57-63. http://doi.org/10.46536/jpumhs/2024/14.01.495

Abstract

BACKGROUND: Polycythemia Vera (PV) is a chronic myeloproliferative neoplasm, the condition arise when
the excessive number of red blood cell produced within the bone marrow due to the mutation occurs at the myeloid
stem cells in protein named JAK2 kinase particularly within JH2 domain. A somatic point mutation occurs in the
JAK2 gene on chromosome 9 due to the transversion of G-C to T-A which develops the replacement of valine by
phenylalanine at codon 617. According to the WHO JAK2 V617F is responsible for about 95% of PV cases. The
annual incidents for Polycythemia Vera are 1-3 individual per 100,000 populations. The aim of the study was to
investigate the prevalence of the JAK2 gene mutation by using Real-Time PCR among the suspected PV patients
in Khyber Pakhtunkhwa, providing visions into the frequency of this mutation within the province. METHODS:
A total of 300 blood samples were collected from suspected PV patients and various screening test were conducted
to confirm the diagnosis, including: Hematologic parameters such as Red blood cell count (RBCs), Hemoglobin
level (HB), Hematocrit (HCT) and total leukocyte count (TLC) were measured. Peripheral blood smear were
examined to detect the abnormalities in the morphology of red blood cell, differential eosinophil and basophil
counts and presence of blast as well. Bone marrow aspirate and biopsy samples were obtained and examined to
assess cellularity, panmyelosis, megakaryocyte morphology, erythroid and myeloid series cells, bone marrow
eosinophilia, bone marrow iron stores, and reticulin fibrosis grade. Genomic DNA was extracted from the
collected blood samples using the MN kit. Real-Time PCR analysis was then performed to detect the JAK2 V617F
mutation, a hallmark of PV. This molecular analysis provided confirmation of PV diagnosis and enabled precise
identification of individuals carrying the JAK2 mutation. RESULTS: Among the suspected patients, 211 were
confirmed positive through CBC, 144 through special smear, and 102 through bone marrow examination. RealTime PCR analysis of DNA samples revealed the existence of the JAK2 V617F mutation in approximately 97
(95%) of the confirmed PV patients. CONCLUSION: In our study, we consider Real-Time PCR as a sensitive
and accurate method for detecting the JAK2 V617F mutation in Polycythemic (PV) patients based on its
demonstrated performance in previous studies and its widespread adoption in clinical laboratories. The high
prevalence of this mutation, detected in nearly 95% of PV cases, highlights its significance in the pathogenesis of
the disease within the Khyber Pakhtunkhwa population.