title: Genotyping the HFE gene by melting point analysis with the LightCycler system: Pros and cons.
abstract: The assay that combines rapid-cycle PCR with allele-specific fluorescent probe melting profiles performed on the Roche Diagnostics LightCycler is commonly employed for genotyping the HFE gene. We report three illustrative cases of the pros and cons of this method. In two cases, atypical melting curves allows the identification of new DNA substitutions in the HFE gene, whereas, in the third case, a typical melting curve of c.845G>A mutation (C282Y) homozygosity overlooks a nucleotide change and promotes misdiagnosis of HH.
authors.affiliation: Servei de Genètica, Hospital de Sant Pau, Avda. Pare Claret 167, 08025 Barcelona, Spain.
authors.forename: V
authors.initials: V
authors.lastname: Bach
authors.forename: M J
authors.initials: MJ
authors.lastname: Barceló
authors.forename: A
authors.initials: A
authors.lastname: Altés
authors.forename: A
authors.initials: A
authors.lastname: Remacha
authors.forename: J
authors.initials: J
authors.lastname: Félez
authors.forename: M
authors.initials: M
authors.lastname: Baiget
mesh_terms: D000328:Adult; D004252:DNA Mutational Analysis / Q000295:instrumentation / Q000379:methods*; D003951:Diagnostic Errors; D005838:Genotype; D006432:Hemochromatosis / Q000175:diagnosis*; D000071020:Hemochromatosis Protein; D015395:Histocompatibility Antigens Class I / Q000235:genetics*; D006358:Hot Temperature; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D008875:Middle Aged; D044367:Phase Transition; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: C103673:HFE protein, human; D000071020:Hemochromatosis Protein; D015395:Histocompatibility Antigens Class I; D008565:Membrane Proteins