20Jan 2017

Utility of Amplification Refractory Mutation System PCR for the detection of JAK2 mutation in Chronic Myeloproliferative Disorders.

  • Prince Fahd Bin Sultan Research chair, Division of cancer Molecular Genetics, Department of Medical Lab Technology, Faculty of Applied Medical Sciences, University of Tabuk, Kingdom of Saudi Arabia.
  • Deparment of Clinical Pathology, King Fahad Special Hospital, Ministry of Health Kingdom of Saudi Arabia.
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Detection of the JAK2 V617F mutation is helpful in differential diagnosis, prognosis, and predication of therapeutic response in CMPD disorders. Because the mutation can be present in a small proportion of granulocytic populations in MPD patients, a highly sensitive detection method is required. The study was aimed to optimize ARMS-PCR for the detection of JAK2V617F mutations .This study was conducted at Prince Fahd Bin Sultan Research Chair .Fifty subjects were registered from the King Khaled Hospital of Tabuk -Northwestern part of Saudi Arabia among which 25 newly diagnosed MPD patients and 25 healthy controls. DNA was extracted done using DNA extraction Kit from Qiagen .ARMS was optimized in which a a primers were designed in such a way that it is able to discriminate among templates that differ by a single nucleotide residue in one tube reaction.Both cases and controls were tested for JAK2 mutations by ARMS-PCR. Demographic data, spleen size, hemoglobin level, white blood cell and platelet counts were recorded. Independent sample t-test was used to study the correlation of JAK2 mutations with age, haemoglobin, blood counts and spleen size. Fisher\'s exact test was applied to compare disease progression in mutation positive and negative cases. This study was conducted on 25 newly diagnosed MPD patients among which 17 were polycythemia vera,5 Essential thrombocytopenia and 3 primary myelofibrosis and 25 healthy controls. Both cases and controls were tested for JAK2 mutations by ARMS-PCR. The JAK2 mutation was detected in 17/25 samples among which 13 were Polycythemia Vera, 02 from Essential thrombocythemia, 01 from Primary myelofibrosis. The JAK2 mutation was not detected in any of the healthy controls by ARMS-PCR. The main advantage of ARMS was that the amplification step and the diagnostic steps were combined, in that the presence of an amplified product indicates the presence of a particular allele and vice versa. For routine diagnosis, this characteristic of ARMS means that it is a very time-efficient method.It was concluded that the ARMS-PCR assay optimized allows the discrimination between homozygous and heterozygous individuals with the JAK2 V617F mutation and has a key role in acting as a reliable screening test for the presence or absence of the mutation in individuals with MPDs. The test will be potentially useful to follow minimal residual disease when targeted therapies against JAK2 kinase become available. The test is therefore a simple, fast, and inexpensive procedure that does not entail any special equipment other than a thermocycler.


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[Rashid Mir, Abu-Duhier FM, Jamsheed J, Fawzia Sharaf & Nabil Burrow. (2017); Utility of Amplification Refractory Mutation System PCR for the detection of JAK2 mutation in Chronic Myeloproliferative Disorders. Int. J. of Adv. Res. 5 (1). 829-837] (ISSN 2320-5407). www.journalijar.com


Rashid Mir


DOI:


Article DOI: 10.21474/IJAR01/2822       DOI URL: http://dx.doi.org/10.21474/IJAR01/2822


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