Ann Hematol (2005) 84: 103–105 DOI 10.1007/s00277-004-0966-4 ORIGINAL ARTICLE Paola E. Leone . Patricia Giménez . Juan Carlos Collantes . César Paz-y-Miño Analysis of HFE gene mutations (C282Y, H63D, and S65C) in the Ecuadorian population Received: 5 July 2004 / Accepted: 1 October 2004 / Published online: 29 October 2004 # Springer-Verlag 2004 Abstract Type 1 hemochromatosis is a disorder of iron metabolism mostly related to the HFE gene mutations. In the present study, we performed a mutation analysis to determine the frequencies of the HFE gene mutations (C282Y, H63D, and S65C) in DNA samples of 100 healthy Ecuadorian individuals. We used the polymerase chain reaction (PCR) to amplify exons 2 and 4 of the HFE gene and then the restriction fragment length polymor- phism (RFLP) method to detect the mutations. The results revealed that the mutations in the normal Ecuadorian population have frequencies of 0.0, 0.035, and 0.04 for C282Y, H63D, and S65C, respectively. We also searched for these mutations in 12 hemochromatosis patients, and the frequencies that we found were 0.0 for C282Y, 0.167 for H63D, and 0.042 for S65C. We found differences [using the chi-square (χ 2 ) test] in the frequency of the H63D mutation between the control group and the group of hemochromatosis patients (p<0.01). This suggests that in Ecuador, type 1 hemochromatosis is more influenced by the H63D mutation than the other two mutations that we analyzed. Given that in a Caucasian population hereditary hemochromatosis is mostly related to the C282Y mutation, it is possible that the findings for the Ecuadorian population are due to geographical differences between the populations. Keywords Ecuador . HFE gene . C282Y . H63D . S65C . Type 1 hemochromatosis Introduction Type 1 hemochromatosis is an autosomal recessive disorder of iron regulation that results in excessive intestinal absorption of iron. This failure of the iron metabolism leads to diverse illnesses such as diabetes mellitus, cardiomyopathy, and liver disease due to a progressive iron loading in parenchymal cells of important organs [1]. In 1996, Feder et al. [2] found a candidate gene for hemochromatosis (HFE), which is localized in human chromosome 6p21.3. This new gene showed two muta- tions in hemochromatosis patients: C282Y (c.845G>A/p. C282Y) and H63D (c.187C>G/p.H63D) [3]. The C282Y mutation has been found in homozygosity in 60–100% of individuals of European descent who present type 1 hemochromatosis, whereas the H63D mutation has a frequency of approximately 16% in the European popu- lation [2, 3]. A third mutation in the HFE gene has been reported in hemochromatosis patients and controls, the S65C (c.193A>T/p.S65C) [4, 5]; arguably this mutation is responsible for a mild form of the disorder [5, 6]. Nonetheless, it is reported that in some cases hemo- chromatosis is not only related to the HFE gene mutations. These interesting findings arise from the observations that within large families showing a high incidence of iron overload disorders, nonaffected members could share the same haplotype of the HFE gene with affected ones. This confirms the hypothesis that at least two genes must be involved in the development of hemochromatosis [7]. In this regard, other genes that have shown association with different types of hemochromatosis have been reported (e.g., HJV, HAMP, and TfR2). These suggest the hetero- geneous feature of the disease [8]. The aim of this study was to determine the status of these three mutations of the HFE gene (C282Y, H63D, and S65C) in Ecuador. This research is of great interest to us because the Ecuadorian population shows a different genetic background compared to other populations [9, 10]. Since there is no report on these allelic frequencies in Ecuador, it was very important to determine them and P. E. Leone . C. Paz-y-Miño Unidad de Genética, Facultad de Medicina, Pontificia Universidad Católica del Ecuador, Quito, Ecuador P. E. Leone (*) . P. Giménez . J. C. Collantes . C. Paz-y-Miño Laboratorio de Genética Molecular y Citogenética Humana, Escuela de Biología, Pontificia Universidad Católica del Ecuador, Quito, Ecuador e-mail: pleone@puce.edu.ec Tel.: +593-2-2991688 Fax: +593-2-2991688