7 research outputs found
Expansion of a BCR-ABL clone in a JAK2 V617F myeloproliferative neoplasm treated by ruxolitinib.
International audienc
Syndrome myéloprolifératif avec polyglobulie et translocation t(6;8)(q27;p11) : cas clinique et revue de la littérature
International audienc
Interlaboratory development and validation of a HRM method applied to the detection of JAK2 exon 12 mutations in polycythemia vera patients.
BACKGROUND: Myeloproliferative disorders are characterized by clonal expansion of normal mature blood cells. Acquired mutations giving rise to constitutive activation of the JAK2 tyrosine kinase has been shown to be present in the majority of patients. Since the demonstration that the V617F mutation in the exon 14 of the JAK2 gene is present in about 90% of patients with Polycythemia Vera (PV), the detection of this mutation has become a key tool for the diagnosis of these patients. More recently, additional mutations in the exon 12 of the JAK2 gene have been described in 5 to 10% of the patients with erythrocytosis. According to the updated WHO criteria the presence of these mutations should be looked for in PV patients with no JAK2 V617F mutation. Reliable and accurate methods dedicated to the detection of these highly variable mutations are therefore necessary. METHODS/FINDINGS: For these reasons we have defined the conditions of a High Resolution DNA Melting curve analysis (HRM) method able to detect JAK2 exon 12 mutations. After having validated that the method was able to detect mutated patients, we have verified that it gave reproducible results in repeated experiments, on DNA extracted from either total blood or purified granulocytes. This HRM assay was further validated using 8 samples bearing different mutant sequences in 4 different laboratories, on 3 different instruments. CONCLUSION: The assay we have developed is thus a valid method, adapted to routine detection of JAK2 exon 12 mutations with highly reproducible results
Clinical and biological data of 8 patients with exon 12 mutations tested in 4 laboratories (RCM: Red cell mass. ND: Not done).
<p>Clinical and biological data of 8 patients with exon 12 mutations tested in 4 laboratories (RCM: Red cell mass. ND: Not done).</p
HRM profiles on 3 different instruments for 2 patients.
<p>Total blood DNA from 2 patients was analysed on 3 different instruments together with non mutated DNA. Arrows show mutant DNA.</p
HRM analysis.
<p>HRM profiles of 7 patients known to harbor JAK2 exon 12 mutations compared to 7 controls. Analysis of total blood DNA on the ABI7500 fast instrument.</p