title: Novel method for molecular detection of the two common hereditary hemochromatosis mutations.
abstract: We describe a novel molecular screening technique for hereditary hemochromatosis through which HFE genotypes at codon positions 282 and 63 are simultaneously detected. The technique combines multiplex PCR and denaturing high-performance liquid chromatography (DHPLC) and allows automated high-throughput analysis. We used this method to genotype 43 previously characterized anonymous DNA specimens in blinded fashion and found multiplex PCR/DHPLC 100% accurate when compared with PCR/restriction enzyme digestion, yet far more efficient.
authors.affiliation: Children's National Medical Center, Washington, DC 20010, USA.
authors.forename: S G
authors.initials: SG
authors.lastname: Kaler
authors.forename: J M
authors.initials: JM
authors.lastname: Devaney
authors.forename: E L
authors.initials: EL
authors.lastname: Pettit
authors.forename: R
authors.initials: R
authors.lastname: Kirshman
authors.forename: M A
authors.initials: MA
authors.lastname: Marino
mesh_terms: D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003062:Codon; D017931:DNA Primers; D006580:Genetic Carrier Screening; D006680:HLA Antigens / Q000235:genetics*; D006432:Hemochromatosis / Q000175:diagnosis / Q000235:genetics*; D000071020:Hemochromatosis Protein; D015395:Histocompatibility Antigens Class I / Q000235:genetics*; D006801:Humans; D008565:Membrane Proteins; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D003062:Codon; D017931:DNA Primers; C103673:HFE protein, human; D006680:HLA Antigens; D000071020:Hemochromatosis Protein; D015395:Histocompatibility Antigens Class I; D008565:Membrane Proteins