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Paper Description


The Development of a Simple and Rapid Diagnostic Test for Gain Loss of Function Defects in Fibroblast Growth Factor-23 (FGF-23).


Authors: Ramadan Ahmed, Al-Ghamdi Manal, Shawar Said
Abstract: FGF-23 is a recently discovered hormone that regulates the homeostasis of inorganic phosphate (pi) and vitamin D. Mutations that generate a loss of function in FGF-23 cause a commonly prevalent disorder in the middle East and Africa Known as familial tumoral calcinosis (FTC). Gain of function mutation in the hormone cause Autosomal dominant hypophospatemic rickets (ADHR). To date, the only method to diagnose defects in FGF-23 is DNA sequence analysis. In this investigation, we report a rapid and simple method to diagnose gain or loss of function in FQF-23 using amplification refractory mutation system (ARMS)-PCR. We employed in vitro site –directed mutagenesis, ARMS-PCR and DNA sequence analysis to produce a rapid diagnostic test for FTC and ADHR due to defects in FGF-23. Results: In vitro site –directed mutagenesis was used to create seven different mutations in FGF-23 that cause FTC and two other mutations that cause ADHR. The mutated sequences were confirmed by DNA sequence analysis. We designed ARMS specific primers to identify each of the mutations and we optimized the conditions required to run the procedure. To validate our diagnostic method, we tested two large Middle East clans clinically diagnosed with FTC. Our results indicate the successful use of this method to diagnose defects in FGF-23. This diagnostic method simplifies and accelerates the detection of FTC and ADHR due to mutations in FGF-23 in an area where the DNA sequence analysis is costly and not commonly found. The high prevalence of FTC is most likely due to high rates of consanguinity in this area . For this reason, the development of rapid and simple methods of diagnosis for genetic diseases is important in prenatal genetic screening and to identify carriers of the disorder and conduct proper counseling to those individuals. We are working on a high resolution melt (HRM) diagnostic method to detect other genes associated ith FTC and can supplement ARMS-PCR.
Published in: The HUGO J;5(1):285, 270-271
URL: http://www.cags.org.ae/4thpahgc. Pdf & http://www.springerlink.com
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