Simple Multiplex PCR for the Simultaneous Detection of the C282Y and H63D Hemochromatosis (HFE) Gene Mutations

Hemochromatosis is a common autosomal recessive disorder of iron metabolism occurring with a prevalence of 0.2–0.5% in Caucasian populations (1)(2)(3)(4)(5)(6). The disease is characterized by the excessive accumulation of dietary iron and a progressive rise in body iron stores, which may lead to serious clinical consequences, including cirrhosis, cardiac failure, diabetes, arthritis, and hepatocellular carcinoma. Treatment involves removal of the iron burden by regular venesection and leads to a normal life expectancy if implemented before the development of cirrhosis (7). Thus early detection and treatment are critically important. Recent identification of a hemochromatosis gene, ( HFE , initially termed HLA-H ) by Feder et al.(8) allows for early genetic diagnosis and greatly simplifies the screening of a family once affected individuals have been identified. The HFE gene encodes a protein similar in structure to MHC class I-type molecules (9) that interacts with the transferrin receptor to regulate iron absorption (10). Two mutations have been detected in the HFE gene. Most individuals with hemochromatosis (80–100%) are homozygous for the missense mutation C282Y. In addition, a small number of compound heterozygotes (heterozygous for both C282Y and H63D) may develop clinical iron overload (11)). Homozygosity for H63D is not clearly associated with hemochromatosis. A high prevalence of asymptomatic carriers of C282Y (13.2%) and H63D (24.3%) …