Heat-transfer-based detection of SNPs in the PAH gene of PKU patients

Natalie Vanden Bon,1 Bart van Grinsven,2 Mohammed Sharif Murib,2 Weng Siang Yeap,2 Ken Haenen,2,3 Ward De Ceuninck,2,3 Patrick Wagner,2,3 Marcel Ameloot,1 Veronique Vermeeren,1 Luc Michiels11Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium; 2Institute for Materials Research, H...

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Autores principales:	Vanden Bon N, van Grinsven B, Murib MS, Yeap WS, Haenen K, De Ceuninck W, Wagner P, Ameloot M, Vermeeren V, Michiels L
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2014
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/2563b333efd84d9785d29b9c2386d86f
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spelling	oai:doaj.org-article:2563b333efd84d9785d29b9c2386d86f2021-12-02T05:39:46ZHeat-transfer-based detection of SNPs in the PAH gene of PKU patients1178-2013https://doaj.org/article/2563b333efd84d9785d29b9c2386d86f2014-03-01T00:00:00Zhttp://www.dovepress.com/heat-transfer-based-detection-of-snps-in-the-pah-gene-of-pku-patients-a16261https://doaj.org/toc/1178-2013 Natalie Vanden Bon,1 Bart van Grinsven,2 Mohammed Sharif Murib,2 Weng Siang Yeap,2 Ken Haenen,2,3 Ward De Ceuninck,2,3 Patrick Wagner,2,3 Marcel Ameloot,1 Veronique Vermeeren,1 Luc Michiels11Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium; 2Institute for Materials Research, Hasselt University, Diepenbeek, Belgium; 3IMOMEC, Diepenbeek, BelgiumAbstract: 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 (Rth) 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 Rth change could be observed during the thermal denaturation of these DNA fragments after hybridization to their complementary counterpart. A clear Rth shift during the denaturation of exon 5, exon 9, and exon 12 dsDNA was observed, corresponding to lengths of up to 123 bp. Finally, Rth 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.Keywords: mutation detection, heat-transfer resistance, melting temperature, nanocrystalline diamond, persistence lengthVanden Bon Nvan Grinsven BMurib MSYeap WSHaenen KDe Ceuninck WWagner PAmeloot MVermeeren VMichiels LDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2014, Iss Issue 1, Pp 1629-1640 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Vanden Bon N van Grinsven B Murib MS Yeap WS Haenen K De Ceuninck W Wagner P Ameloot M Vermeeren V Michiels L Heat-transfer-based detection of SNPs in the PAH gene of PKU patients
description	Natalie Vanden Bon,1 Bart van Grinsven,2 Mohammed Sharif Murib,2 Weng Siang Yeap,2 Ken Haenen,2,3 Ward De Ceuninck,2,3 Patrick Wagner,2,3 Marcel Ameloot,1 Veronique Vermeeren,1 Luc Michiels11Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium; 2Institute for Materials Research, Hasselt University, Diepenbeek, Belgium; 3IMOMEC, Diepenbeek, BelgiumAbstract: 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 (Rth) 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 Rth change could be observed during the thermal denaturation of these DNA fragments after hybridization to their complementary counterpart. A clear Rth shift during the denaturation of exon 5, exon 9, and exon 12 dsDNA was observed, corresponding to lengths of up to 123 bp. Finally, Rth 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.Keywords: mutation detection, heat-transfer resistance, melting temperature, nanocrystalline diamond, persistence length
format	article
author	Vanden Bon N van Grinsven B Murib MS Yeap WS Haenen K De Ceuninck W Wagner P Ameloot M Vermeeren V Michiels L
author_facet	Vanden Bon N van Grinsven B Murib MS Yeap WS Haenen K De Ceuninck W Wagner P Ameloot M Vermeeren V Michiels L
author_sort	Vanden Bon N
title	Heat-transfer-based detection of SNPs in the PAH gene of PKU patients
title_short	Heat-transfer-based detection of SNPs in the PAH gene of PKU patients
title_full	Heat-transfer-based detection of SNPs in the PAH gene of PKU patients
title_fullStr	Heat-transfer-based detection of SNPs in the PAH gene of PKU patients
title_full_unstemmed	Heat-transfer-based detection of SNPs in the PAH gene of PKU patients
title_sort	heat-transfer-based detection of snps in the pah gene of pku patients
publisher	Dove Medical Press
publishDate	2014
url	https://doaj.org/article/2563b333efd84d9785d29b9c2386d86f
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Heat-transfer-based detection of SNPs in the PAH gene of PKU patients

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