A reverse-hybridization assay for the rapid and simultaneous detection of nine HFE gene mutations

Hereditary hemochromatosis (HH) is a very common autosomal recessive disord er of iron metabolism and frequently associated with mutations in the HFE g ene. Molecular genetic testing for HFE mutations is considered valuable for carrier identification, as well as for early diagnosis of the disease, all owing simple treat. ment by phlebotomy and normal survival of patients. We have developed a reverse-hybridization assay for the routine diagnosis of e ight previously described and one novel (E168Q) HFE point mutations The tes t is based on multiplex DNA amplification and ready-to-use membrane teststr ips, which contain oliganucleotide probes for each wild-type and mutated al lele immobilized as an array of parallel lines. The procedure is rapid and accessible to automation on commercially available equipment, and by adding new probes the teststrip can easily be adapted to cover an increasing numb er of mutations.