20Jan 2017

Optimization of allele specific PCR (AS-PCR) for the early detection of FLT3 (D835Y) mutation in Acute Myeloid Leukemia patients at Tabuk, Saudi Arabia.

  • Prince Fahd Bin Sultan Research chair, Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, University of Tabuk, Kingdom of Saudi Arabia.
  • King Fahd specialist Hospital, Ministry of Health, Tabuk, Saudi Arabia.
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Activating mutations in FLT3 occur in ~30% of adult AML, primarily consisting of internal tandem duplication (ITD) mutations (~25%) and point mutations in the tyrosine kinase domain (~7%), at the activation loop (residue D835). The early detection of FLT3- D835Y in AML patients is clinically important for AML patients in this small region of Saudi Arabia especially for determining drug responses, prognosis, and selection of therapy., The present study was conducted on 47 specimens among which 37 were from leukemia patients (5 were newly diagnosed AML, 7 ALL and 25 MPD ) and 10 healthy controls. Genomic DNA from blood was isolated using DNA QIAamp DNA Blood Mini Kit from Qiagen and stored at -200C until PCR. The Allele Specific-PCR based assay for FLT3 (D835Y) mutation was optimized in the Lab .The mutation was detected in 2/5 cases of AML, 0/25 of MPD and 0/5 of ALL. It was observed that patients with AML possessed significantly higher frequency of FLT3 D835 mutation (2/5) than other leukemia cases and healthy controls. The D835Y Flt3-TKD mutation was not detected in any of the healthy controls (0/10). It was concluded that allele specific PCR technique permits the direct detection of FLT3 c.2503G>T (D835Y) point mutation in AML patients and acts as useful genetic technique that is very valuable for molecular diagnosis, prognosis, drug response, and predisposition to AML patients. In addition, the test was simple, fast, and inexpensive procedure that does not require any special equipment other than a thermo cycler.


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[Abu-Duhier FM, Rashid Mir, Jamsheed Javid, Fawzia Sharaf and Nabil Burrow. (2017); Optimization of allele specific PCR (AS-PCR) for the early detection of FLT3 (D835Y) mutation in Acute Myeloid Leukemia patients at Tabuk, Saudi Arabia. Int. J. of Adv. Res. 5 (Jan). 817-822] (ISSN 2320-5407). www.journalijar.com


Dr Faisal AbuDuhier
Prince Fahd Bin Sultan Research chair, Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, University of Tabuk, Kingdom of Saudi Arabia.

DOI:


Article DOI: 10.21474/IJAR01/2819      
DOI URL: http://dx.doi.org/10.21474/IJAR01/2819