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Visual screening for JAK2V617F mutation by a disposable dipstick

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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.

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Authors and Affiliations

  1. Laboratory of Analytical Chemistry, Department of Chemistry, Athens University, 15771, Athens, Greece

    Jessica K. Konstantou, Alexandra C. Iliadi & Penelope C. Ioannou

  2. Department of Chemistry, University of Patras, 26500, Patras, Greece

    Theodore K. Christopoulos

  3. Foundation for Research and Technology Hellas, Institute of Chemical Engineering and High Temperature Chemical Processes (FORTH/ICE-HT), 26504, Patras, Greece

    Theodore K. Christopoulos

  4. Department of Clinical Haematology, General Hospital of Athens ‘G. Gennimatas’, 11527, Athens, Greece

    Nikolaos I. Anagnostopoulos

  5. Laboratory of Medical Genetics, Athens University, 11527, Athens, Greece

    Emmanuel Kanavakis & Jan Traeger-Synodinos

Authors
  1. Jessica K. Konstantou
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  2. Alexandra C. Iliadi
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  3. Penelope C. Ioannou
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  4. Theodore K. Christopoulos
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  5. Nikolaos I. Anagnostopoulos
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  6. Emmanuel Kanavakis
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  7. Jan Traeger-Synodinos
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Corresponding author

Correspondence to Penelope C. Ioannou.

Additional information

J.K.K. and A.C.I contributed equally to this work.

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Konstantou, J.K., Iliadi, A.C., Ioannou, P.C. et al. Visual screening for JAK2V617F mutation by a disposable dipstick. Anal Bioanal Chem 397, 1911–1916 (2010). https://doi.org/10.1007/s00216-010-3747-z

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  • DOI: https://doi.org/10.1007/s00216-010-3747-z

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