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

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• 作者：Alline Didone^a ; Luciana Nardinelli^a ; Mariana Marchiani^a ; Antonio Roberto Lancha Ruiz^a ; Ariel Lais de Lima Costa^a ; Ismael Severino Lima^a ; Nathalia Moreira Santos^a ; Sabri Saeed Sanabani^b ; Israel Bendit^a ; ^{isbendit@usp.br" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Myeloproliferative ; JAK2 V617F ; Mutation ; Wild type ; Screening
• 刊名：Practical Laboratory Medicine
• 出版年：2016
• 出版时间：1 April 2016
• 年：2016
• 卷：4
• 期：Complete
• 页码：30-37
• 全文大小：705 K

文摘

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.