Genotyping the HFE gene by melting point analysis with the LightCycler system: Pros and cons

V Bach; M J Barceló; A Altés; A Remacha; J Félez; M Baiget

doi:10.1016/j.bcmd.2005.12.030

Genotyping the HFE gene by melting point analysis with the LightCycler system: Pros and cons

Blood Cells Mol Dis. 2006 Mar-Apr;36(2):288-91. doi: 10.1016/j.bcmd.2005.12.030. Epub 2006 Mar 2.

Authors

V Bach¹, M J Barceló, A Altés, A Remacha, J Félez, M Baiget

Affiliation

¹ Servei de Genètica, Hospital de Sant Pau, Avda. Pare Claret 167, 08025 Barcelona, Spain.

PMID: 16515868
DOI: 10.1016/j.bcmd.2005.12.030

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.

Publication types

Case Reports
Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Adult
DNA Mutational Analysis / instrumentation
DNA Mutational Analysis / methods*
Diagnostic Errors
Genotype
Hemochromatosis / diagnosis*
Hemochromatosis Protein
Histocompatibility Antigens Class I / genetics*
Hot Temperature
Humans
Male
Membrane Proteins / genetics*
Middle Aged
Phase Transition*
Polymerase Chain Reaction
Sensitivity and Specificity

Substances

HFE protein, human
Hemochromatosis Protein
Histocompatibility Antigens Class I
Membrane Proteins