A sensitive and reliable semi-quantitative real-time PCR assay to detect JAK2 V617F in blood

摘要: Diagnosis of the myeloproliferative disorders, polycythemia vera (PV), essential thrombocythemia (ET) and idiopathic myelofibrosis (IMF) is difficult due to lack diagnostic markers. Recently, acquisition a mutation in Janus kinase 2 (JAK2) gene by hemopoietic cells has been described as genetic defect underlying disorders. The leads constitutive activation JAK2, tyrosine involved cytokine receptor signalling. Because clinical importance this (JAK2 V617F) diagnosing disorders its relevance for disease progression, we developed semi-quantitative real-time PCR test detect JAK2 V617F. With assay, quantities down 0.8% V617F amongst wild-type DNA could reliably be detected. For quantification purposes, low intra- inter-assay variabilities ensure good reproducibility assay. Thus qPCR assay here quick, robust, simple more sensitive than direct sequencing, RFLP, ARMS other methods published so far We therefore believe that will contribute early diagnosis management, especially when JAK2-specific inhibitors have become available therapeutic use.