Evaluation of High Resolution Melting Technique for Detection of JAK2-V617F Mutation in Formalin-Fixed Paraffin-Embedded Specimen from Myeloproliferative Neoplasm Cases

Authors

  • Mohammad Reza Abdullahi Abdul Saboor
  • Nor Zamzila Abdullah
  • Rosmawati Ismail
  • Naznin Muhammad
  • Norlelawati A. Talib

DOI:

https://doi.org/10.31436/imjm.v17i1.847

Abstract

Introduction: Myeloproliferative neoplasm (MPN) is a group of myeloid disorder which lead to erythrocytosis, thrombocytosis and leukocytosis. MPN with BCR-ABL positive is chronic myeloid leukemia (CML) while BCR-ABL negative MPN includes polycythemia 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 cross-sectional study. Forty FFPE marrow specimens were retrieved from the year 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 3 out of 4 in PV, 0/2 in ET, 4/4 in PMF, 4/10 in non-classified MPN and 1/20 in CML. 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.

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Published

2018-12-14

How to Cite

Abdul Saboor, M. R. A., Abdullah, N. Z. ., Ismail, R., Muhammad, N. ., & A. Talib, N. . (2018). Evaluation of High Resolution Melting Technique for Detection of JAK2-V617F Mutation in Formalin-Fixed Paraffin-Embedded Specimen from Myeloproliferative Neoplasm Cases. IIUM Medical Journal Malaysia, 17(1). https://doi.org/10.31436/imjm.v17i1.847

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