title: Validation of fluorescence-labeled artificial nonhuman sequences for single-strand conformation polymorphism mutation detection in familial hypercholesterolemia.
abstract: We developed a two-in-one, polymerase chain reaction (PCR)-based method with a specific amplification step and a universal amplification step in one tube to screen for the presence of DNA variations. The method relies on fluorescence-labeled artificial nonhuman sequences for mutation detection. To document utility, we applied this method as a high-throughput capillary single-strand conformation polymorphism screening system to identify 30 mutations in the low-density lipoprotein receptor gene. The sensitivity of mutant allele detection compared to wild-type allele detection was 93%. We conclude that the "two-in-one PCR" is sensitive, simple, and cost effective.
authors.affiliation: Franz Volhard Clinic, HELIOS Kliniken-Berlin, 13122 Berlin, Germany.
authors.forename: Atakan
authors.initials: A
authors.lastname: Aydin
authors.forename: Friedrich C
authors.initials: FC
authors.lastname: Luft
authors.forename: Sylvia
authors.initials: S
authors.lastname: Bähring
mesh_terms: D000818:Animals; D004252:DNA Mutational Analysis; D017931:DNA Primers; D019075:Electrophoresis, Capillary; D005456:Fluorescent Dyes; D014644:Genetic Variation; D006938:Hyperlipoproteinemia Type II / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D011973:Receptors, LDL / Q000235:genetics*; D015203:Reproducibility of Results; D013194:Staining and Labeling
chemical_list: D017931:DNA Primers; D005456:Fluorescent Dyes; D011973:Receptors, LDL