Simultaneous detection of HFE C282Y, H63D and S65C mutations associated with type 1 haemochromatosis using a multiplex luminex bead assay.

摘要: Type 1 hereditary haemochromatosis (HH) is a common genetic disorder in Caucasoids resulting from mutations the HFE gene. Routine diagnostic testing for type HH involves genotyping two of these described mutations, C282Y and H63D. In some cases typing third mutation, S65C also performed. Several techniques have been reported include polymerase chain reaction (PCR)-sequence-specific primers (SSP), PCR-restriction fragment length polymorphism (RFLP), PCR-sequence-specific oligonucleotide probe (SSOP), real-time PCR followed by melting curve analysis TaqMan assay. The aim this study was to develop an alternative method both conventional PCR/TaqMan assay detect all three single using Luminex technology. DNA controls known genotypes (n = 109) were used evaluate approach. These selected represent possible (wild type, mutant, heterozygous) each mutation. Subsequently, blind samples 100) validate method. This new then compared with current (in-house PCR-SSP assay). Comparison obtained those previously in-house showed 100% concordance no discrepancies observed. Allelic frequency C282Y, H63D 22%, 16% 2%, respectively. We report here high-throughput, accurate robust multiplex luminex bead routine clinical