Abstract | BACKGROUND: METHODS: Using a tri-primer ARMS-PCR, in vitro site-directed mutagenesis and DNA sequencing, we developed, verified and validated a rapid and reliable diagnostic test for the ten mutations in FGF23. RESULTS: We generated a test for all ten mutations and confirmed each test by DNA sequencing. We increased the specificity of the test by introducing a mismatch at position -2 in the 3'-terminus of the reverse primer of the normal and the mutant sequences. Finally, using DNA sequencing, we validated the technique for FGF23/S129F substitution by testing samples from 80 individuals from two unrelated Arab families harboring HFTC. CONCLUSIONS: This inexpensive and specific method could be adopted where DNA sequencing is not available or affordable.
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Authors | Ahmad R Ramadan, Said M Shawar, Manal A Alghamdi |
Journal | Hormone research in paediatrics
(Horm Res Paediatr)
Vol. 86
Issue 1
Pg. 45-52
( 2016)
ISSN: 1663-2826 [Electronic] Switzerland |
PMID | 27355663
(Publication Type: Clinical Trial, Journal Article)
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Copyright | © 2016 S. Karger AG, Basel. |
Chemical References |
- FGF23 protein, human
- Fibroblast Growth Factors
- Fibroblast Growth Factor-23
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Topics |
- Amino Acid Substitution
- Calcinosis
(genetics)
- DNA Mutational Analysis
(methods)
- Female
- Fibroblast Growth Factor-23
- Fibroblast Growth Factors
(genetics)
- Humans
- Hyperostosis, Cortical, Congenital
(genetics)
- Hyperphosphatemia
(genetics)
- Male
- Mutation, Missense
- Polymerase Chain Reaction
(methods)
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