MutaGEL® HFE PCR KIT
Hemochromatosis (C282Y, H63D, S65C)
INTENDED USE
The MutaGEL® HFE PCR kit (hemochromatosis) allows for the genotyping of the three most abundant mutations of the human HFE gene on chromosome 6: in codon 63 amino acid change His to Asp = DNA base change C to G, in codon 65 Ser to Cys = A to T and in codon 282 Cys to Tyr = G to A.
INTRODUCTION
The most frequent recessive heritable disease of Homo sapiens is the chronically upregulated iron uptake of the organ tissues called idiopathic hemochromatosis, which becomes symptomatic with severe liver failure and diabetes in the second part of live and which is accompanied mostly by homozygosity of DNA mutations of the HFE gene in its codon 282 (282Tyr/Tyr) or combined heterozygozity 63Asp/282Tyr or 63Asp63Asp and very rarely 65Tyr together with others. The pathogenic effect of the mutated HFE proteins is today discussed as being the iron receptor block at crypt cells of the duodenum or loss of HFE's function as sensor of the iron concentration in the blood (hepcidin hypothesis). The most accurate test for confirmation of the disease is the iron saturation of transferrin. Early therapy (bleeding) of the severe and common disease is easy and therefore prophylactic measurement of transferrin saturation and HFE genotype both as confirmational test and for screening of the relatives of a patient for identification of undetected disease is helpful. Early symtoms are „the three As“ arthralgia + asthenia + elevated aminotransferases.
PRINCIPLE OF THE TEST
MutaGEL® HFE PCR kit
The MutaGEL® HFE PCR kit (hemochromatosis) consists of three test sets: 1 (double) allele specific kit for typing of the most abundant mutation in codon 282 and two RFLP kits for the typing of the mutations in codon 63 and 65. Therefore in the 282 set of reagents the both possible DNA bases are derected by two allele-specific PCRs in parallel and the two possible DNA bases in codon 63 and 65 each are detected by regional PCRs, followed by restriction enzyme digestion, the enzymes cutting in the case of existent mutations and not cutting in the case of normal sequences.
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