BACKGROUND: Two mutations in @entity14398 , @entity49310 ( @entity535 substitution @entity14399 ) and @entity49311 ( @entity14397 ), are associated with @entity9595 . We developed and validated a novel method, linked linear amplification (LLA), for detection of these two mutations. METHODS: Two segments of @entity14398 were amplified by a multiplex LLA reaction that generated biotinylated LLA products. Aliquots of the multiplex LLA reaction were captured in microwells by hybridization to immobilized allele-specific oligonucleotides (ASOs). One wild-type and one mutant ASO represented the DNA sequence at each of the two mutation sites. Hybridization was detected by a streptavidin-horseradish peroxidase-based colorimetric method. Genotypes obtained by LLA and PCR-restriction fragment length polymorphism (PCR-RFLP) methods for 320 individuals were compared. RESULTS: The amplified samples included the following genotypes as determined by PCR-RFLP: wild-type 282 and 63 codons (n = 105), @entity14399 homozygous mutant (n = 54), @entity14399 heterozygous (n = 52), @entity14397 homozygous mutant (n = 17), @entity14397 heterozygous (n = 59), and compound @entity14397 and @entity14399 heterozygous mutant (n = 33). There was complete concordance between the results obtained by LLA and those obtained by PCR-RFLP analysis. The presence of another @entity14398 mutation, @entity69138 (encoding @entity31397 ), did not interfere with genotyping at codon 63. CONCLUSIONS: LLA provides a reliable method to detect the common mutations in @entity14398 that cause @entity9595 .

Linked linear amplification for simultaneous analysis of the two most common XXXX mutations.

@entity14398 :: ('3077', 'Gene') :: ['HFE'] @entity69138 :: ('c|SUB|A|193|T', 'DNAMutation') :: ['A193T'] @entity49310 :: ('c|SUB|G|845|A', 'DNAMutation') :: ['G845A'] @entity535 :: ('CHEBI:33704', 'Chemical') :: ['amino acid'] @entity14399 :: ('p|SUB|C|282|Y', 'ProteinMutation') :: ['C282Y'] @entity14397 :: ('p|SUB|H|63|D', 'ProteinMutation') :: ['H63D'] @entity49311 :: ('c|SUB|C|187|G', 'DNAMutation') :: ['C187G'] @entity9595 :: ('MESH:D006432', 'Disease') :: ['hereditary hemochromatosis'] @entity31397 :: ('p|SUB|S|65|C', 'ProteinMutation') :: ['S65C']
@entity9595 :: (MESH:D006432,Disease) :: ['hemochromatosis']
