Mutation analysis of the hamartin gene using denaturing high performance liquid chromatography

Denaturing high performance liquid chromatography (DHPLC) is a novel high‐capacity technique for gene mutation scanning. We have assessed the sensitivity and specificity of this method for analysis of the full coding sequence of the hamartin (TSC1) gene in 20 tuberous sclerosis patients, whose TSC1...

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Bibliographic Details
Published in:Human mutation 2000-11, Vol.16 (5), p.417-421
Main Authors: Bénit, Paule, Kara-Mostefa, Ali, Berthelon, Monique, Sengmany, Kamchan, Munnich, Arnold, Bonnefont, Jean-Paul
Format: Article
Language:English
Subjects:
Chromatography, High Pressure Liquid - methods
denaturing high-performance liquid chromatography
DHPLC
DNA - blood
DNA Mutational Analysis
Female
Genes, Tumor Suppressor - genetics
hamartin
Humans
Male
mutation scanning
Nucleic Acid Denaturation - genetics
Polymorphism, Genetic - genetics
Proteins - genetics
TSC1
tuberous sclerosis
Tuberous Sclerosis - genetics
Tumor Suppressor Proteins
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Summary:Denaturing high performance liquid chromatography (DHPLC) is a novel high‐capacity technique for gene mutation scanning. We have assessed the sensitivity and specificity of this method for analysis of the full coding sequence of the hamartin (TSC1) gene in 20 tuberous sclerosis patients, whose TSC1 genes previously had been studied by single strand conformation polymorphism analysis and protein truncation assay. All eight sequence variants previously identified were adequately detected by DHPLC. Additionally, this approach picked up three polymorphisms, one of which (IVS13‐55 C>G) was hitherto unreported, therefore serving as proof of principle for this technique. Thus, DHPLC appears to be a highly sensitive method with advantages in terms of flexibility, fragments size analysis, cost and time and labor sparing, compared to classical approaches of mutation scanning. Hum Mutat 16:417–421, 2000. © 2000 Wiley‐Liss, Inc.
ISSN:1059-7794
1098-1004
DOI:10.1002/1098-1004(200011)16:5<417::AID-HUMU6>3.0.CO;2-4