title: Development of a quantitative real-time polymerase chain reaction assay for the detection of the JAK2 V617F mutation.
abstract: Achieving a specific diagnosis of polycythemia vera (PV) and other myeloproliferative disorders (MPDs) is often costly and complex. However, the recent identification of a V617F mutation in the JH2 domain of the JAK2 gene in a high proportion of patients suffering from MPDs may provide confirmation of a diagnosis. This is an acquired mutation and, as such, may only be present in a small number of cells within a sample. There is therefore a clinical need for highly sensitive detection techniques. We have developed a sensitive real-time polymerase chain reaction (PCR)-based approach for both detection and quantification of the JAK2 V671F mutation load, which allows determination of mutation status without the need for prior purification of granulocytes. We have performed a comparison of this assay with two previously published detection methods. Although an amplification refractory mutation system (ARMS) was shown to be slightly superior in terms of sensitivity, our real-time PCR method provides the potential for quantification of the JAK2 V617F mutation, having potential future applications in the monitoring of minimal residual disease or predicting outcome of disease severity.
authors.affiliation: Molecular Genetics Department, Royal Devon and Exeter NHS Foundation Trust, Barrack Rd., Exeter, UK EX2 5DW.
authors.forename: Elizabeth C
authors.initials: EC
authors.lastname: Wolstencroft
authors.forename: Katy
authors.initials: K
authors.lastname: Hanlon
authors.forename: Lorna W
authors.initials: LW
authors.lastname: Harries
authors.forename: Graham R
authors.initials: GR
authors.lastname: Standen
authors.forename: Alexander
authors.initials: A
authors.lastname: Sternberg
authors.forename: Sian
authors.initials: S
authors.lastname: Ellard
mesh_terms: D017931:DNA Primers; D006801:Humans; D053614:Janus Kinase 2 / Q000235:genetics*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D020125:Mutation, Missense / Q000235:genetics*; D011087:Polycythemia Vera / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; C507924:JAK2 protein, human; D053614:Janus Kinase 2