title: A rapid PCR-SSP assay for the hemochromatosis-associated Tyr250Stop mutation in the TFR2 gene.
abstract: Several genes associated with hemochromatosis and primary iron overload have been identified. Mutations in the HFE gene have been detected in 60-100% of hemochromatosis patients of northern, central, and western European descent, although the frequencies of these mutations vary among racial and ethnic groups. Recently, a mutation in the gene encoding transferrin receptor-2 (exon 6, nucleotide 750 C --> G; Y250X) was detected by a PCR-restriction fragment length polymorphism (RFLP) method in Sicilians with hemochromatosis. We describe a modification of the original assay in which the sequence-specific priming PCR assay does not require the use of restriction endonuclease. The modified assay is robust and cost-efficient, and may be more useful for large-scale population studies because it can be performed rapidly on DNA extracted from buccal swabs.
authors.affiliation: Department of Microbiology, University of Alabama at Birmingham, 35294, USA.
authors.forename: C A
authors.initials: CA
authors.lastname: Rivers
authors.forename: J C
authors.initials: JC
authors.lastname: Barton
authors.forename: R T
authors.initials: RT
authors.lastname: Acton
mesh_terms: D000328:Adult; D000349:Africa / Q000208:ethnology; D000407:Alabama / Q000453:epidemiology; D002610:Cheek; D003062:Codon / Q000235:genetics; D018389:Codon, Nonsense; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004252:DNA Mutational Analysis / Q000191:economics / Q000379:methods*; D004534:Egypt / Q000208:ethnology; D005006:Ethnicity / Q000235:genetics; D005060:Europe / Q000208:ethnology; D005260:Female; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000191:economics / Q000379:methods*; D006432:Hemochromatosis / Q000453:epidemiology / Q000208:ethnology / Q000235:genetics*; D006801:Humans; D007558:Italy / Q000208:ethnology; D007861:Lebanon / Q000208:ethnology; D008297:Male; D009061:Mouth Mucosa / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D011990:Receptors, Transferrin / Q000235:genetics*; D013997:Time Factors
chemical_list: D003062:Codon; D018389:Codon, Nonsense; D011990:Receptors, Transferrin; C503640:TFR2 protein, human; D004247:DNA