Novel method for molecular detection of the two common hereditary hemochromatosis mutations.
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.
Kaler
S G
SG
Children's National Medical Center, Washington, DC 20010, USA.
Devaney
J M
JM
Pettit
E L
EL
Kirshman
R
R
Marino
M A
MA
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
D003062:Codon; D017931:DNA Primers; C103673:HFE protein, human; D006680:HLA Antigens; D000071020:Hemochromatosis Protein; D015395:Histocompatibility Antigens Class I; D008565:Membrane Proteins