Population screening for hemochromatosis by PCR using sequence-specific primers

Over 90% of patients with hemochromatosis in the United Kingdom are homozyg ous for the C282Y mutation on the HFE gene. The Centers for Disease Control (CDC) in the United States has recommended that adults should be screened for HFE mutations to identify susceptible individuals before onset of disea se. The aim of this study was to evaluate the polymerase chain reaction usi ng sequence-specific primers (PCR-SSP) as a method of large-scale populatio n screening for the common HFE gene mutations, H63D and C282Y, A total of 1 0,583 consenting blood donors were tested using nonautomated procedures. Th ree alleles, termed HFE-1, -2, and -3, mere detected with phenotype frequen cies of 94.56%, 28.33%, and 15.79%, respectively, and gene frequencies of 0 .76421, 0.15342, and 0.08237, respectively. All donors identified as homozy gous for the C282Y mutation or heterozygous for both the H63D and C282Y mut ations mere confirmed by heterduplex analysis and/or PCR-SSP, The number of technical failures that affected the identification of donors homozygous f or the C282Y mutation was 390 giving an overall repeat rate 3.7%, although this fell to 1% over the last quarter of the study, This study demonstrates that PCR-SSP may be used for large-scale population screening for the C282 Y genotype associated with hemochromatosis.