Quantitative Determination of JAK2 V617F by TaqMan: An Absolute Measure of Averaged Copies per Cell That May Be Associated with the Different Types of Myeloproliferative Disorders

摘要: We report a novel TaqMan assay for JAK2 V617F that measures averaged copies per cell in absolute terms, as opposed to ratio of mutant wild-type alleles. Measurements were obtained by comparing the signal generated test samples set external plasmid standards containing sequence interest. Specificity was demonstrated above 36 cycles amplification, and endpoint titration experiments indicated sensitivity down 0.05% clinical dilutions. The measured linearly over wide logarithmic range exhibited good reproducibility. Combination this with another method determining number allowed identification 14 cases myeloproliferative disease greater than two cell. Mutational frequency 68% among polycythemia vera (n = 44), 59% 37) essential thrombocythemia 46% 13) idiopathic myelofibrosis. Levels mutation significantly higher compared (P 0.0005) correlated following jointly significant variables at diagnosis: PRV-1, hemoglobin, white count, neutrophil red using multiple regression analyses 0.015). This should be useful assessing relationship gene dose phenotype possibly monitoring therapy.