Evaluation of high resolution melting technique for detection of JAK2-V617F mutation in formalin-fixed paraffin-embedded specimen from myeloproliferative neoplasm cases

• Main Authors: Abdullahi, Mohammad Reza, Abdullah, Nor Zamzila, Ismail, Rosmawati, Muhammad, Siti Aeshah @ Naznin, A.Talib, Norlelawati
• Format: Article
• Language: English; English
• Published: Kulliyyah (Faculty) of Medicine, International Islamic University Malaysia 2019
• Subjects: R Medicine (General); RB Pathology
• Online Access: http://irep.iium.edu.my/76120/1/76120_Evaluation%20of%20high%20resolution%20melting%20technique%20for%20detection%20of%20JAK2-V617F%20mutation%20in%20formalin-fixed%20paraffin-embedded%20specimen%20from%20myeloproliferative%20neoplasm%20cases.pdf; http://irep.iium.edu.my/76120/2/76120_Evaluation%20of%20high%20resolution%20melting%20technique%20for%20detection%20of%20JAK2-V617F%20mutation%20in%20formalin-fixed%20paraffin-embedded%20specimen%20from%20myeloproliferative%20neoplasm%20case_SCOPUS.pdf; http://irep.iium.edu.my/76120/; http://iiumedic.net/imjm/v1/download/volume_18_no_2/Pages-from-IMJMVol18No2-15.pdf

Introduction: Myeloproliferative neoplasm (MPN) is a group of myeloid disorders which leads to erythrocytosis, thrombocytosis and leucocytosis. MPN with BCR-ABL positive is chronic myeloid leukaemia (CML) while BCR-ABL negative MPN includes polycythaemia Vera (PV), essential thrombocytemia (ET) and primary myelofibrosis (PMF). One of the major criteria for diagnosis of BCR-ABL negative MPN is the presence of JAK2-V617F mutation which is positive in 95% of PV and around 60% of ET and MF. Beside peripheral blood specimen, formalin-fixed paraffin-embedded (FFPE) marrow specimen can be used for detection of this mutation. Unfortunately, FFPE produces low quality DNA that put a challenge for successful amplification of DNA. We aimed to evaluate the utility of High Resolution Melting (HRM) analysis for detection of JAK2-V617F mutation in FFPE specimen from MPN cases. Materials and Methods: This study is a descriptive crosssectional study. Forty FFPE marrow specimens were retrieved from the years 2014-2016. Bio-Rad Precision Melt Analysis software was used for analysis of HRM data. Allele-specific PCR was done for validation of results. Positive samples were subjected to Sanger sequencing. Results: JAK2-V617F mutation was positive in 13 out of 40 MPN cases. Level of agreement between HRM and AS-PCR was 97.5%. Conclusion: HRM is a rapid and powerful diagnostic assay which is suitable for detection of JAK2-V617F mutation in FFPE marrow specimen.