title: A tetra-primer polymerase chain reaction approach for the detection of JAK2 V617F mutation.
abstract: Recently, an acquired somatic point mutation (p.V617F) in a highly conserved residue of the pseudokinase domain of the JAK2 tyrosine kinase was shown to be associated with myeloproliferative disorders. Because of the clinical importance of this mutation in diagnosing myeloproliferative disorders and its relevance for disease progression, we have developed a tetra-primer polymerase chain reaction (PCR) assay to detect JAK2 p.V617F. Titration studies showed that the assay could reliably detect one copy of the mutant allele in a mix of 50 wild-type alleles suggesting that the lower detection limit of this assay is estimated to be 2%. This study demonstrates that genotyping and quantifying of the JAK2 V617F mutation can be performed by tetra-primer PCR using both freshly isolated and formalin-fixed tissues. Our tetra-primer PCR assay is sensitive, low-cost, and easy to use method for the detection of JAK2 p.V617F, which could be used even in low-tech laboratories.
authors.affiliation: Burc Molecular Diagnostic Laboratory, Vali Konagi Cad, Istanbul, Turkey.
authors.forename: Vedat
authors.initials: V
authors.lastname: Koksal
authors.forename: Ozdal
authors.initials: O
authors.lastname: Etlik
authors.forename: S Tugba
authors.initials: ST
authors.lastname: Arican-Baris
authors.forename: Ibrahim
authors.initials: I
authors.lastname: Baris
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000097:blood; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D006801:Humans; D053614:Janus Kinase 2 / Q000097:blood / Q000235:genetics*; D008875:Middle Aged; D017354:Point Mutation; D011087:Polycythemia Vera / Q000175:diagnosis / Q000201:enzymology / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D015415:Biomarkers; D017931:DNA Primers; D053614:Janus Kinase 2