title: Detection of the JAK2 V617F mutation by LightCycler PCR and probe dissociation analysis.
abstract: A point mutation in the JAK2 gene, a member of the tyrosine kinase family, was recently identified and shown to be associated with several myeloproliferative disorders. Several studies identified the same JAK2 point mutation (1,849G>T), resulting in the substitution of a valine to phenylalanine at codon 617 (V617F). We developed a simple and sensitive method to detect this mutation via polymerase chain reaction and probe dissociation analysis using the LightCycler platform, and we compared this method to existing restriction fragment-length polymorphism, direct sequencing, and amplification refractory mutation system methods. We found that the LightCycler method offered advantages of speed, reliability, and more straightforward interpretation over the restriction fragment-length polymorphism and sequencing approaches.
authors.affiliation: Molecular Pathology Laboratory, L235, 300 Pasteur Dr., Stanford University School of Medicine, CA 94305, USA.
authors.forename: Marla
authors.initials: M
authors.lastname: Lay
authors.forename: Rajan
authors.initials: R
authors.lastname: Mariappan
authors.forename: Jason
authors.initials: J
authors.lastname: Gotlib
authors.forename: Lisa
authors.initials: L
authors.lastname: Dietz
authors.forename: Siby
authors.initials: S
authors.lastname: Sebastian
authors.forename: Iris
authors.initials: I
authors.lastname: Schrijver
authors.forename: James L
authors.initials: JL
authors.lastname: Zehnder
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods* / Q000592:standards; D006801:Humans; D053614:Janus Kinase 2; D015336:Molecular Probe Techniques; D009196:Myeloproliferative Disorders / Q000175:diagnosis / Q000235:genetics; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012984:Software; D044366:Transition Temperature
chemical_list: D011518:Proto-Oncogene Proteins; D011505:Protein-Tyrosine Kinases; C507924:JAK2 protein, human; D053614:Janus Kinase 2