title: Multiplex detection and genotyping of point mutations involved in charcot-marie-tooth disease using a hairpin microarray-based assay.
abstract: We previously developed a highly specific method for detecting SNPs with a microarray-based system using stem-loop probes. In this paper we demonstrate that coupling a multiplexing procedure with our microarray method is possible for the simultaneous detection and genotyping of four point mutations, in three different genes, involved in Charcot-Marie-Tooth disease. DNA from healthy individuals and patients was amplified, labeled with Cy3 by multiplex PCR; and hybridized to microarrays. Spot signal intensities were 18 to 74 times greater for perfect matches than for mismatched target sequences differing by a single nucleotide (discrimination ratio) for "homozygous" DNA from healthy individuals. "Heterozygous" mutant DNA samples gave signal intensity ratios close to 1 at the positions of the mutations as expected. Genotyping by this method was therefore reliable. This system now combines the principle of highly specific genotyping based on stem-loop structure probes with the advantages of multiplex analysis.
authors: Department of Biochemistry and Molecular Genetics, University of Limoges, 87025 Limoges, France.
authors: Yasser
authors: Y
authors: Baaj
authors: Corinne
authors: C
authors: Magdelaine
authors: Virginie
authors: V
authors: Ubertelli
authors: Christophe
authors: C
authors: Valat
authors: Yoanne
authors: Y
authors: Mousseau
authors: Hao
authors: H
authors: Qiu
authors: Benoît
authors: B
authors: Funalot
authors: Jean-Michel
authors: JM
authors: Vallat
authors: Franck G
authors: FG
authors: Sturtz