General information
The SALSA MLPA
Probemix P255-B1 ALDOB-FBP1 is a
research use only (RUO) assay for the detection of deletions or duplications in the
ALDOB and
FBP1, which are associated with Hereditary fructose intolerance.
Hereditary fructose intolerance (HFI) is a rare recessive inherited disorder of carbohydrate metabolism, caused by catalytic deficiency of the aldolase B enzyme (ALDOB). The ALDOB enzyme plays a key role in glycolysis and gluconeogenesis and, in mammals, is preferentially expressed in the liver. HFI patients manifest hypoglycemia, lactic acidosis, and gastrointestinal symptoms, such as severe abdominal pain and recurrent vomiting after consuming fructose-containing foods. HFI usually presents in infancy at the time of weaning, when fructose is added to the diet. Persistent ingestion of fructose and related sugars (such as sucrose and sorbitol) can lead to severe liver and kidney damage, seizures, coma, and risk of death.
Mutations in the aldolase B gene (
ALDOB) cause hereditary fructose intolerance (HFI). The protein expressed by this gene catalyses the cleavage of fructose-1-phosphate to form dihydroxyacetone phosphate and D-glyceraldehyde.
Hereditary fructose-1,6-diphosphatase deficiency is another form of fructose intolerance and exhibits similar signs and symptoms to HFI. This condition is inherited in an autosomal recessive way and caused by a deficiency in fructose-1,6-bisphosphatase 1 (FBP1), an enzyme which catalyses the hydrolysis of fructose 1,6-bisphosphate to fructose 6-phosphate and inorganic phosphate.
The
ALDOB gene (9 exons), spans ~15.2 kb of genomic DNA and is located on chromosome 9q31.1, 101.4 Mb from p-telomere. The
FBP1 gene (8 exons), spans ~37.1 kb of genomic DNA and is located on chromosome 9q22.32, 96.4 Mb from p-telomere.
More information is available at
https://www.ncbi.nlm.nih.gov/books/NBK333439/.
More information is available at
https://www.ncbi.nlm.nih.gov/books/NBK550349/.
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 P255-B1 contains 32 MLPA probes with amplification products between 163 and 445 nucleotides (nt). P255-B1 contains probes for each exon of the
ALDOB gene, including two mutation-specific probes (A149P / c.448G>C; A174D / c.524C>A). In addition, one flanking probe targeting the
TGFBR1 gene located 2.3 Mb downstream of
ALDOB is included. Furthermore, this probemix contains probes for each exon of the
FBP1 gene. In addition, twelve 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.
SALSA Binning DNA SD030
The SD030 Binning DNA provided with this probemix can be used for binning of all probes including the two mutation specific probes (
ALDOB / exon 5 probe 08669-L08680, mutation c.448G>C and
ALDOB / exon 5 probe 08670-L08682, mutation c.524C>A. SD030 Binning DNA is a mixture of genomic DNA from healthy individuals and plasmid DNA that contains the target sequence detected by the above mentioned probe. Inclusion of one reaction with 5 μl SD030 Binning DNA in initial MLPA experiments is essential as it can be used to aid in data binning of the peak pattern using Coffalyser.Net software. Furthermore, Binning DNA should be included in the experiment whenever changes have been applied to the set-up of the capillary electrophoresis device (e.g. when capillaries have been renewed). Binning DNA should never be used as a reference sample in the MLPA data analysis, neither should it be used in quantification of mutation signal(s). It is strongly advised that all samples tested are extracted with the same method and derived from the same source of tissue. For further details, please consult the SD030 Binning DNA product description, available online:
www.mrcholland.com.
This product is for research use only (RUO).
Sample DNA
Sample DNA developed for this product: