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. 2007;3(6):315-24.
doi: 10.3233/cbm-2007-3605.

Detection of the JAK2V617F mutation by asymmetric PCR and melt curve analysis

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Free article

Detection of the JAK2V617F mutation by asymmetric PCR and melt curve analysis

Bobbie Collett Sutton et al. Cancer Biomark. 2007.
Free article

Abstract

The chronic myeloproliferative disorders (CMPDs) are a heterogeneous group of clonal hematopoietic diseases characterized by production of increased numbers of mature leukocytes, erythrocytes, and/or platelets. Clinically these disorders are often insidious in onset, produce nonspecific thrombotic or hemorrhagic complications, and can be easily confused with a variety of benign, reactive conditions. Thus, confirming a CMPD can be difficult as it is often a diagnosis of exclusion. The recently identified JAK2(V617F) mutation is frequently present in the classic CMPDs polycythemia vera, essential thrombocythemia, and chronic idiopathic myelofibrosis. JAK2(V617F) determination has proven to be a useful diagnostic tool in patients with some clinical features suggestive for a CMPD, and may have benefit as a way to monitor known disease. There are several published molecular assays for the JAK2(V617F) target, of variable sensitivity and technical complexity, many of which are not easily replicated in a typical clinical laboratory. We present a robust, sensitive PCR/melt curve assay for the JAK2(V617F) mutation which uses the widely available Roche LightCycler platform, and is thus applicable to many clinical molecular laboratories.

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