Detection of the JAK2 V617F Mutation by LightCycler PCR and Probe Dissociation Analysis

• Published in: The Journal of molecular diagnostics : JMD Vol. 8; no. 3; pp. 330 - 334
• Main Authors: Lay, Marla, Mariappan, Rajan, Gotlib, Jason, Dietz, Lisa, Sebastian, Siby, Schrijver, Iris, Zehnder, James L.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2006; ASIP; American Society for Investigative Pathology
• Subjects: 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; Regular; Restriction Mapping; Sensitivity and Specificity; Sequence Analysis, DNA; Software; Transition Temperature
• ISSN: 1525-1578; 1943-7811
• DOI: 10.2353/jmoldx.2006.050130

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 (1849G>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.