Avtor/Urednik		Cukjati, M; Vaupotič, T; Rupreht, R; Čurin-Šerbec, V
Naslov		Prevalence of H63D, S65C and C282Y hereditary hemochromatosis gene mutations in Slovenian population by an improved high-throughput genotyping assay
Tip		članek
Vir		BMC Med Genet
Vol. in št.		Letnik 8
Leto izdaje		2007
Obseg		str. 1-9, 69
Jezik		eng
Abstrakt		Background: Hereditary hemochromatosis (HH) is a common genetic disease characterized by excessive iron overload that leads to multi-organ failure. Although the most prevalent genotype in HH is homozygosity for C282Y mutation of the HFE gene, two additional mutations, H63D and S65C, appear to be associated with a milder form of HH. The aim of this study was to develop a high-throughput assay for HFE mutations screening based on TaqMan technology and to determine the frequencies of HFE mutations in the Slovenian population. Methods: Altogether, 1282 randomly selected blood donors from different Slovenian regions and 21 HH patients were analyzed for the presence of HFE mutations by an in-house developed real-time PCR assay based on TaqMan technology using shorter non-interfering fluorescent single nucleotide polymorphism (SNP)-specific MGB probes. The assay was validated by RFLP analysis and DNA sequencing. Results: The genotyping assay of the H63D, S65C and C282Y mutations in the HFE gene, based on TaqMan technology proved to be fast, reliable, with a high-throughput capability and 100% concordant with genotypes obtained by RFLP and DNA sequencing. The observed frequency of C282Y homozygotes in the group of HH patients was only 48%, others were of the heterogeneous HFE genotype. Among 1282 blood donors tested, the observed H63D, S65C and C282Y allele frequency were 12.8% (95% confidence interval (CI) 11.5-14.2%), 1.8% (95% CI 1.4-2.5%) and 3.6% (95% CI 3.0-4.5%), respectively. Approximately 33% of the tested subjects had at least one of the three HH mutations, and 1% of them were C282Y homozygotes or compound heterozygotes C282Y/H63D or C282Y/S65C, presenting an increased risk for iron overload disease. A significant variation in H63D allele frequency was observed for one of the Slovenian regions. (Abstract truncated at 2000 characters)
Deskriptorji		MUTATION ADULT AGED CHI-SQUARE DISTRIBUTION FLUORESCENT DYES GENE FREQUENCY GENOTYPE HEMOCHROMATOSIS HISTOCOMPATIBILITY ANTIGENS CLASS I MEMBRANE PROTEINS POLYMERASE CHAIN REACTION POLYMORPHISM, RESTRICTION FRAGMENT LENGTH PREVALENCE SEQUENCE ANALYSIS, DNA SLOVENIA