Detection of the JAK2 V617F mutation by LightCycler PCR and probe dissociation analysis

J Mol Diagn. 2006 Jul;8(3):330-4. doi: 10.2353/jmoldx.2006.050130.

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.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • DNA Mutational Analysis / methods*
  • DNA Mutational Analysis / standards
  • Humans
  • Janus Kinase 2
  • Molecular Probe Techniques
  • Myeloproliferative Disorders / diagnosis
  • Myeloproliferative Disorders / genetics
  • Point Mutation
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Restriction Fragment Length*
  • Protein-Tyrosine Kinases / genetics*
  • Proto-Oncogene Proteins / genetics*
  • Restriction Mapping
  • Sensitivity and Specificity
  • Sequence Analysis, DNA
  • Software
  • Transition Temperature

Substances

  • Proto-Oncogene Proteins
  • Protein-Tyrosine Kinases
  • JAK2 protein, human
  • Janus Kinase 2