An unusual melting curve profile in LightCycler multiplex genotyping of the hemochromatosis H63D/C282Y gene mutations.
OBJECTIVE
Real time polymerase chain reaction followed by melting curve analysis using hybridization probes has become an important tool in routine diagnosis of the HFE mutations, which are associated with hereditary hemochromatosis.


METHODS
We used the LightCycler technology for simultaneous detection of the H63D and C282Y mutations of the HFE gene in patients with a higher prevalence for hemochromatosis.


RESULTS
In our cohort we identified two siblings with a variant pattern of the HFE-LightCycler melting profiles preventing allelic discrimination.


CONCLUSIONS
As a consequence, in these patients DNA sequencing or RFLP analysis is necessary to unequivocally assign the correct HFE genotype.
Tag
C G
CG
Institute of Clinical Chemistry and Pathobiochemistry-Central Laboratory, RWTH-University Hospital, Pauwelsstr. 30, D-52074 Aachen, Germany.
Gressner
A M
AM
Weiskirchen
R
R
D019943:Amino Acid Substitution; D016477:Artifacts; D015331:Cohort Studies; D004252:DNA Mutational Analysis; D005787:Gene Frequency; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D005858:Germany / Q000453:epidemiology; D006432:Hemochromatosis / Q000175:diagnosis* / Q000453:epidemiology / Q000235:genetics*; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D009154:Mutation; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D015995:Prevalence; D012016:Reference Values