title: Heat-transfer-based detection of SNPs in the PAH gene of PKU patients.
abstract: Conventional neonatal diagnosis of phenylketonuria is based on the presence of abnormal levels of phenylalanine in the blood. However, for carrier detection and prenatal diagnosis, direct detection of disease-correlated mutations is needed. To speed up and simplify mutation screening in genes, new technologies are developed. In this study, a heat-transfer method is evaluated as a mutation-detection technology in entire exons of the phenylalanine hydroxylase (PAH) gene. This method is based on the change in heat-transfer resistance (R(th)) upon thermal denaturation of dsDNA (double-stranded DNA) on nanocrystalline diamond. First, ssDNA (single-stranded DNA) fragments that span the size range of the PAH exons were successfully immobilized on nanocrystalline diamond. Next, it was studied whether an R(th) change could be observed during the thermal denaturation of these DNA fragments after hybridization to their complementary counterpart. A clear R(th) shift during the denaturation of exon 5, exon 9, and exon 12 dsDNA was observed, corresponding to lengths of up to 123 bp. Finally, R(th) was shown to detect prevalent single-nucleotide polymorphisms, c.473G>A (R158Q), c.932T>C (p.L311P), and c.1222C>T (R408W), correlated with phenylketonuria, displaying an effect related to the different melting temperatures of homoduplexes and heteroduplexes.
authors.affiliation: Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium.
authors.forename: Natalie
authors.initials: N
authors.lastname: Vanden Bon
authors.affiliation: Institute for Materials Research, Hasselt University, Diepenbeek, Belgium.
authors.forename: Bart
authors.initials: B
authors.lastname: van Grinsven
authors.affiliation: Institute for Materials Research, Hasselt University, Diepenbeek, Belgium.
authors.forename: Mohammed Sharif
authors.initials: MS
authors.lastname: Murib
authors.affiliation: Institute for Materials Research, Hasselt University, Diepenbeek, Belgium.
authors.forename: Weng Siang
authors.initials: WS
authors.lastname: Yeap
authors.affiliation: Institute for Materials Research, Hasselt University, Diepenbeek, Belgium ; IMOMEC, Diepenbeek, Belgium.
authors.forename: Ken
authors.initials: K
authors.lastname: Haenen
authors.affiliation: Institute for Materials Research, Hasselt University, Diepenbeek, Belgium ; IMOMEC, Diepenbeek, Belgium.
authors.forename: Ward
authors.initials: W
authors.lastname: De Ceuninck
authors.affiliation: Institute for Materials Research, Hasselt University, Diepenbeek, Belgium ; IMOMEC, Diepenbeek, Belgium.
authors.forename: Patrick
authors.initials: P
authors.lastname: Wagner
authors.affiliation: Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium.
authors.forename: Marcel
authors.initials: M
authors.lastname: Ameloot
authors.affiliation: Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium.
authors.forename: Veronique
authors.initials: V
authors.lastname: Vermeeren
authors.affiliation: Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium.
authors.forename: Luc
authors.initials: L
authors.lastname: Michiels
mesh_terms: D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000379:methods*; D005819:Genetic Markers / Q000235:genetics; D020022:Genetic Predisposition to Disease / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D010651:Phenylalanine Hydroxylase / Q000235:genetics*; D010661:Phenylketonurias / Q000175:diagnosis* / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013817:Thermography / Q000379:methods*; D044366:Transition Temperature
chemical_list: D005819:Genetic Markers; D010651:Phenylalanine Hydroxylase
keywords: heat-transfer resistance; melting temperature; mutation detection; nanocrystalline diamond; persistence length
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