The thrombopoietin receptor W515L and W515k mutations detection in patients with essential thrombocythemia

• Published in: Zdravniški vestnik (Ljubljana, Slovenia : 1992) Slovenia : 1992), 2012-12, Vol.81 (SUPL II)
• Main Authors: Pajic, Tadej, Kovacic, Leonida, Mlakar, Uros
• Format: Article
• Language: English
• ISSN: 1318-0347; 1581-0024; 1581-0224

Background: The aim of the study was to investigate the frequency of the thrombopoietin receptor (MPL) W515L and W515K mutations in our cohort of essential thrombocythemia (ET) patients negative for the somatic JAK2 V617F mutation, and to compare the results to those published in the literature. Methods: Seventy-seven ET patients (59 female, 18 male), median age 48 (range from 18 to 89 years) and negative for JAK2 V617F mutation were included in the study. Granulocytes were isolated from patients' venous blood samples by Ficoll density gradient centrifugation followed by red blood cell lysis procedure. DNA was isolated from granulocytes by QIAamp DNA Mini Kit from Qiagen (USA). The detection of MPL W515L/K mutations was carried out by test designed by Ipsogen (France) using the fluorescence-based quantitative real-time PCR (qPCR) and allelic discrimination method. Results: MPL W515L and MPL W515K mutations in our cohort of patients with ET and negative for the somatic JAK2 V617F mutation was found in 4 % (N = 3) and 2 % (N = 1), respectively. Both mutations were found in 5 % of ET patients included in the study (N = 77). Conclusions: We have found out that the frequency of MPL W515L/K mutation was similar to those published in the literature and was 5 %. The MPL W515L mutation frequency was higher than MPL W515K. The assay is suitable for the routine detection of MPL W515L/K mutations in patients with ET and negative for the somatic JAK2 V617F mutation.