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

Abdullahi, Mohammad Reza and Abdullah, Nor Zamzila and Ismail, Rosmawati and Muhammad, Siti Aeshah @ Naznin and A.Talib, Norlelawati (2019) Evaluation of high resolution melting technique for detection of JAK2-V617F mutation in formalin-fixed paraffin-embedded specimen from myeloproliferative neoplasm cases. International Medical Journal Malaysia, 18 (2). pp. 95-100. E-ISSN 1823-4631

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Abstract

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.

Item Type:	Article (Review)
Additional Information:	3746/76120
Uncontrolled Keywords:	High Resolution Melting (HRM), Allele-Specific PCR (AS-PCR), JAK2 Mutation, Myeloproliferative neoplasm (MPN), FFPE
Subjects:	R Medicine > R Medicine (General) R Medicine > RB Pathology
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Medicine Kulliyyah of Medicine > Department of Basic Medical Kulliyyah of Medicine > Department of Pathology & Lab Medicine
Depositing User:	Dr Nor Zamzila Abdullah
Date Deposited:	10 Dec 2019 15:04
Last Modified:	06 Apr 2022 10:07
URI:	http://irep.iium.edu.my/id/eprint/76120

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