Comparative study of different methodologies to detect the JAK2 V617F mutation in chronic BCR-ABL1 negative myeloproliferative neoplasms.

Autor:	Didone A; Laboratorio de Biologia Tumoral Disciplina de Hematologia do Hoispital de Clínicas da Faculdade de Medicina da Universidade de Sao Paulo, Brazil., Nardinelli L; Laboratorio de Biologia Tumoral Disciplina de Hematologia do Hoispital de Clínicas da Faculdade de Medicina da Universidade de Sao Paulo, Brazil., Marchiani M; Laboratorio de Biologia Tumoral Disciplina de Hematologia do Hoispital de Clínicas da Faculdade de Medicina da Universidade de Sao Paulo, Brazil., Ruiz ARL; Laboratorio de Biologia Tumoral Disciplina de Hematologia do Hoispital de Clínicas da Faculdade de Medicina da Universidade de Sao Paulo, Brazil., de Lima Costa AL; Laboratorio de Biologia Tumoral Disciplina de Hematologia do Hoispital de Clínicas da Faculdade de Medicina da Universidade de Sao Paulo, Brazil., Lima IS; Laboratorio de Biologia Tumoral Disciplina de Hematologia do Hoispital de Clínicas da Faculdade de Medicina da Universidade de Sao Paulo, Brazil., Santos NM; Laboratorio de Biologia Tumoral Disciplina de Hematologia do Hoispital de Clínicas da Faculdade de Medicina da Universidade de Sao Paulo, Brazil., Sanabani SS; Clinical Laboratory, Department of Pathology, LIM 03, Hospital das Clínicas (HC), School of Medicine, University of São Paulo, São Paulo, Brazil., Bendit I; Laboratorio de Biologia Tumoral Disciplina de Hematologia do Hoispital de Clínicas da Faculdade de Medicina da Universidade de Sao Paulo, Brazil.
Jazyk:	angličtina
Zdroj:	Practical laboratory medicine [Pract Lab Med] 2015 Dec 14; Vol. 4, pp. 30-37. Date of Electronic Publication: 2015 Dec 14 (Print Publication: 2016).
DOI:	10.1016/j.plabm.2015.12.004
Abstrakt:	Objectives: A mutation in the JAK2 gene, V617F, has been identified in several BCR-ABL1 negative myeloproliferative neoplasms (MPN): polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). Defining the presence or absence of this mutation is an essential part of clinical diagnostic algorithms and patient management. Here, we aimed to evaluate the performance of three PCR-based assays: Amplification Refractory Mutation System (ARMS), High-Resolution Melting analysis (HRM), and Sanger direct sequencing, and compare their results with those obtained by a PCR restriction fragment polymorphism assay (PCR-RFLP). Design and Methods: We used blood samples from 136 patients (PV=20; PMF=20; ET=28, and other MPN suspected cases=68). Results: Comparable results were observed among the four assays in patients with PV, PMF, and MPN suspected cases. In patients with a diagnosis of ET, the JAK2 V617F mutation was detected in 67.8% of them by the PCR-ARMS and PCR-HRM assay and in 64% of them by the conventional Sanger sequence approach. The PCR-ARMS and PCR-HRM assays were 100% concordant. With these tests, only one of the 20 patients with ET and one of the three patients with clinically suspected MPN gave different results compared with those obtained by the PCR-RFLP. Conclusions: Our results have demonstrated that the PCR-ARMS and PCR-HRM assays could detect the JAK2 V617F mutation effectively in MPN patients, but PCR-HRM assays are rapid and the most cost-effective procedures.
Databáze:	MEDLINE
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