Myeloid Leukemia

Detecting JAK2 V617F Mutation in MPD 253
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Methods for the Detection of the JAK2 V617F Mutation
in Human Myeloproliferative Disorders
Peter J. Campbell,* Linda M. Scott,* E. Joanna Baxter,
Anthony J. Bench, Anthony R. Green, and Wendy N. Erber
Summary
A single acquired mutation in the JAK2 gene has recently been described in human myeloproliferative
disorders, including most patients with polycythemia vera and about half of those
with essential thrombocythemia and idiopathic myelofibrosis. Reliable and easily implemented
methods for detection of this V617F mutation promise to revolutionize the way these disorders
are diagnosed and classified, and may in the future have implications for targeted therapeutics.
Two polymerase chain reaction-based methods for detection of the mutation are described here.
One method is based on allele-specific amplification of the mutant band, and the other on
elimination of a restriction enzyme recognition sequence by the mutation. Both methods are
significantly more sensitive than conventional sequencing techniques, and could be readily
implemented in a molecular diagnostic laboratory.
Key Words: Myeloproliferative disorder; polycythemia vera; essential thrombocythemia;
idiopathic myelofibrosis; JAK2; mutation.
1. Introduction
The classic myeloproliferative disorders (MPDs) comprise polycythemia
vera (PV), essential thrombocythemia (ET), and idiopathic myelofibrosis (IMF)
(1). They are considered clonal stem cell disorders (2), and result in expansion
of one or more lineages of bone marrow cells, usually with retention of differentiation
capacity. The sine qua non of PV is expansion of the red cell mass,
with most patients also developing granulocytosis and/or thrombocythemia,
whereas ET is generally characterised by an isolated thrombocythemia (3–5).
The major clinical manifestations of these disorders are arterial thrombosis,
venous thrombosis, and a propensity to hemorrhage, but overall life expectancy
* These authors contributed equally to this work.
From: Methods in Molecular Medicine, Vol. 125: Myeloid Leukemia: Methods and Protocols
Edited by: H. Iland, M. Hertzberg, and P. Marlton © Humana Press Inc., Totowa, NJ
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