General information
The SALSA MLPA
Probemix P347 Hemochromatosis is a
research use only (RUO) assay for the detection of deletions or duplications in the
HFE,
TFR2,
HFE2 (HJV),
HAMP, and
SLC40A1 genes, which are associated with hereditary hemochromatosis (HH). This probemix can also be used to detect the presence of the
HFE C282Y point mutation (c.845G>A/p.Cys282Tyr) and it detects the wildtype allele of the
HAMP G71D point mutation (c.212G>A/p.Gly71Asp).
HH is an inherited disorder characterised by progressive iron deposition and tissue injury in multiple organs. HH has been demonstrated to result from mutations in several genes involved in the regulation of iron homeostasis such as
HFE,
TFR2,
HFE2,
HAMP and
SLC40A1. Mutations in these genes act in an autosomal recessive manner, except for
SLC40A1 which manifests in autosomal dominant phenotypes. The most common form of HH is associated with the homozygous C282Y mutation of the
HFE gene which accounts for 83% of the cases (Alexander et al. 2009).
More information is available at
https://www.ncbi.nlm.nih.gov/books/NBK1440/,
https://www.ncbi.nlm.nih.gov/books/NBK1349/, and
https://www.ncbi.nlm.nih.gov/books/NBK1170/.
This SALSA MLPA probemix is not CE/FDA registered for use in diagnostic procedures. Purchase of this product includes a limited license for research purposes.
Probemix content
The SALSA MLPA Probemix P347-A3 Hemochromatosis contains 44 MLPA probes with amplification products between 136 and 481 nucleotides (nt). This includes three probes for the
HFE2 (
HJV) gene, eight probes for the
SLC40A1 gene (one for each exon), ten probes for the
HFE gene (one for each exon), eight probes for the
TFR2 gene and three probes for the
HAMP gene (one for each exon). Furthermore, this probemix contains one probe specific for the
HFE C282Y point mutation, which will only generate a signal when the mutation is present. It also contains one probe specific for the G71D point mutation within the
HAMP gene, which will only generate a signal when the wild-type allele is present. A reduced signal can point towards the presence of the mutation
or a deletion of exon 3. In addition, ten reference probes are included that detect autosomal chromosomal locations. Complete probe sequences and the identity of the genes detected by the reference probes are available online (
www.mrcholland.com).
This probemix contains nine quality control fragments generating amplification products between 64 and 105 nt: four DNA Quantity fragments (Q-fragments), two DNA Denaturation fragments (D-fragments), one Benchmark fragment, and one chromosome X and one chromosome Y-specific fragment. More information on how to interpret observations on these control fragments can be found in the MLPA General Protocol and online at
www.mrcholland.com.