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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, 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: 10 Dec 2019 15:04
URI: http://irep.iium.edu.my/id/eprint/76120

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