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TITLE:

Clinical utility of ARMS-PCR amplification-refractory mutation system for the detection of JAK2 gene variations [JAK2 haplotypes] in myeloproliferative disorders [polycythemia vera, essential thrombocythemia, and primary myelofibrosis in Tabuk.

AUTHORS:

Rashid Mir, Abu-Duhier FM

ABSTRACT:

Background: The germline JAK2 haplotype known as "GGCC or 46/1 haplotype" [haplotypeGGCC_46/1] consists of a combination of single nucleotide polymorphisms [SNPs] mapping in a region of about 250 kb, extending from the JAK2 intron 10 to the Insulin-like 4 [INLS4] gene. Four main SNPs [rs3780367, rs10974944, rs12343867, and rs1159782] generating a "GGCC" combination are more frequently indicated to represent the JAK2 haplotype. These SNPs are inherited together and are frequently associated with the onset of myeloproliferative neoplasms [MPN] positive for both JAK2 V617 and exon 12 mutations. Therefore the aim of our study was to optimize ARMS-PCR system for the detection of JAK2 haplotypes i.e rs3780367T/G and rs12343867 T>C in Myeloid proliferative disorders Methodology: This study was conducted on 88 samples among which 44 were MPD patients and 44 were healthy controls. DNA was extracted by Qiagen Kit and ARMS PCR system was optimized to detect JAK2 haplotype in Myeloid proliferative disorders . Results: The study included 88 specimens among which 44 were newly diagnosed MPD patients .Out of 44 MPD patients 26 [59%] were polycythemia vera, 17 [39%] Essential thrombocytopenia and 1 [2%] primary myelofibrosis and 44 healthy controls. Of 44 consecutive MPD patients, 14 [31.81%] were below or equal to 40 years age and 30 [68%] were above 40 years of age. Of 44 consecutive MPD patients, 30 [38.63%] were males and 27 [61.36%] were females. Among healthy controls 22 [50%] were below or equal to 40 years age and 22 [50%] were above 40 years of age. Of 44 healthy controls, 23 [51%] were males and 21 [49%] were females. ARMS PCR system was optimized to detect JAK2 haplotypes rs3780367 T/G and rs12343867 T>C gene variations in Myeloid proliferative disorders. The technique was successfully optimized by using wild-type or mutant-type primers with matched or one-base mismatched to examine the known SNPS in JAK2 gene. It was indicated that ARMS technique can be used as a potential molecular tool in the detection of potential JAK2 gene variations in MPDs . Conclusion: In our study, we successfully developed the Allele specific ARMS PCR technique for rapid detection of SNPs in JAK2 gene. Taken together, ARMS could be useful for quick and accurate SNPs detection in JAK2 gene in MPD as well as other genetic diseases in undeveloped and developing countries which are in shortage of medical resources and supplies. Keywords: Myeloproliferative disorders [MPD] ARMS-PCR amplification-refractory mutation system ,SNP- Single-nucleotide polymorphism, polycythemia vera [PV], essential thrombocythemia [ET], and primary myelofibrosis [PMF] Corresponding author:

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