Evaluation of high resolution melting technique for detection of JAK2-V617F mutation in formalin-fixed paraffin-embedded specimen from myeloproliferative neoplasm cases
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) an...
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iium-76120 |
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eprints |
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iium-761202019-12-10T07:04:54Z http://irep.iium.edu.my/76120/ 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 Abdullah, Nor Zamzila Ismail, Rosmawati Muhammad, Naznin A.Talib, Norlelawati R Medicine (General) RB Pathology 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. Kulliyyah (Faculty) of Medicine, International Islamic University Malaysia 2019-08 Article PeerReviewed application/pdf en 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 application/pdf en 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 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 http://iiumedic.net/imjm/v1/download/volume_18_no_2/Pages-from-IMJMVol18No2-15.pdf |
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Digital Repository |
| institution_category |
Local University |
| institution |
International Islamic University Malaysia |
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Online Access |
| language |
English English |
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R Medicine (General) RB Pathology |
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R Medicine (General) RB Pathology Abdullahi, Mohammad Reza Abdullah, Nor Zamzila Ismail, Rosmawati Muhammad, Naznin A.Talib, Norlelawati Evaluation of high resolution melting technique for detection of JAK2-V617F mutation in formalin-fixed paraffin-embedded specimen from myeloproliferative neoplasm cases |
| description |
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. |
| format |
Article |
| author |
Abdullahi, Mohammad Reza Abdullah, Nor Zamzila Ismail, Rosmawati Muhammad, Naznin A.Talib, Norlelawati |
| author_facet |
Abdullahi, Mohammad Reza Abdullah, Nor Zamzila Ismail, Rosmawati Muhammad, Naznin A.Talib, Norlelawati |
| author_sort |
Abdullahi, Mohammad Reza |
| title |
Evaluation of high resolution melting technique for detection of JAK2-V617F mutation in formalin-fixed paraffin-embedded specimen from myeloproliferative neoplasm cases |
| title_short |
Evaluation of high resolution melting technique for detection of JAK2-V617F mutation in formalin-fixed paraffin-embedded specimen from myeloproliferative neoplasm cases |
| title_full |
Evaluation of high resolution melting technique for detection of JAK2-V617F mutation in formalin-fixed paraffin-embedded specimen from myeloproliferative neoplasm cases |
| title_fullStr |
Evaluation of high resolution melting technique for detection of JAK2-V617F mutation in formalin-fixed paraffin-embedded specimen from myeloproliferative neoplasm cases |
| title_full_unstemmed |
Evaluation of high resolution melting technique for detection of JAK2-V617F mutation in formalin-fixed paraffin-embedded specimen from myeloproliferative neoplasm cases |
| title_sort |
evaluation of high resolution melting technique for detection of jak2-v617f mutation in formalin-fixed paraffin-embedded specimen from myeloproliferative neoplasm cases |
| publisher |
Kulliyyah (Faculty) of Medicine, International Islamic University Malaysia |
| publishDate |
2019 |
| url |
http://irep.iium.edu.my/76120/ http://irep.iium.edu.my/76120/ 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 |
| first_indexed |
2023-09-18T21:47:36Z |
| last_indexed |
2023-09-18T21:47:36Z |
| _version_ |
1777413554551390208 |