Analytical and Bioanalytical Chemistry

, Volume 397, Issue 5, pp 1911–1916

Visual screening for JAK2V617F mutation by a disposable dipstick

  • Jessica K. Konstantou
  • Alexandra C. Iliadi
  • Penelope C. Ioannou
  • Theodore K. Christopoulos
  • Nikolaos I. Anagnostopoulos
  • Emmanuel Kanavakis
  • Jan Traeger-Synodinos
Original Paper

DOI: 10.1007/s00216-010-3747-z

Cite this article as:
Konstantou, J.K., Iliadi, A.C., Ioannou, P.C. et al. Anal Bioanal Chem (2010) 397: 1911. doi:10.1007/s00216-010-3747-z

Abstract

During the last 5 years, it was discovered that the JAK2V617F somatic mutation is present in virtually all patients with polycythemia vera and a large proportion of patients with essential thrombocythemia, primary myelofibrosis, and refractory anemia with ring sideroblasts and thrombocytosis. As a result, JAK2V617F was incorporated as a new clonal marker in the 2008 revision of the WHO diagnostic criteria. Current methods for JAK2 genotyping include direct sequencing, pyrosequencing, allele-specific PCR with electrophoresis, restriction fragment length polymorphism, real-time PCR, DNA-melting curve analysis, and denaturing HPLC. Some of these methods are labor intensive and time consuming, while the others require specialized costly equipment and reagents. We report a method for direct detection of the JAK2V617F allele by the naked eye using a dipstick test in a dry-reagent format. The method comprises a triprimer PCR combined with visual detection of the products within minutes by the dipstick test. Specialized instrumentation is not involved. The requirements for highly qualified technical personnel are minimized. Because the detection reagents exist in dry form on the dipstick, there is no need for multiple pipetting and incubation steps.

Keywords

JAK2V617F Mutation detection Triprimer PCR Dipstick test 

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Jessica K. Konstantou
    • 1
  • Alexandra C. Iliadi
    • 1
  • Penelope C. Ioannou
    • 1
  • Theodore K. Christopoulos
    • 2
    • 3
  • Nikolaos I. Anagnostopoulos
    • 4
  • Emmanuel Kanavakis
    • 5
  • Jan Traeger-Synodinos
    • 5
  1. 1.Laboratory of Analytical Chemistry, Department of ChemistryAthens UniversityAthensGreece
  2. 2.Department of ChemistryUniversity of PatrasPatrasGreece
  3. 3.Foundation for Research and Technology HellasInstitute of Chemical Engineering and High Temperature Chemical Processes (FORTH/ICE-HT)PatrasGreece
  4. 4.Department of Clinical HaematologyGeneral Hospital of Athens ‘G. Gennimatas’AthensGreece
  5. 5.Laboratory of Medical GeneticsAthens UniversityAthensGreece