LightCycler PCR assay for simultaneous detection of the H63D and S65C mutations in the HFE hemochromatosis gene based on opposite melting temperature shifts

Hereditary hemochromatosis is a common autosomal recessive disorder of iron metabolism, which in its ho-mozygous form occurs in Caucasian populations with a prevalence of 0.2–0.5 % (1, 2). Characteristic of the disease is the excessive accumulation of dietary iron and a pro-gressive rise in iron stores. This may lead to serious clinical consequences, including cirrhosis, cardiac failure, diabetes, arthritis, and hepatocellular carcinoma. Treat-ment involves removal of the iron burden by regular venesection and leads to a normal life expectancy if implemented before the development of cirrhosis (3). Thus, early detection and treatment are critically impor-tant. The recent identification of a hemochromatosis gene (HFE, initially termed HLA-H) by Feder et al. (4) allows early genetic diagnosis and greatly simplifies the screen