The SALSA MLPA Probemix P280 SLC26A4 is a research use only (RUO)
assay for the detection of deletions or duplications in the SLC26A4
gene, which is associated with Pendred syndrome and autosomal recessive non-syndromic deafness (DFNB4) (Cirello et al. 2018). This probemix can also be used to detect the presence of three point mutations: the L236P (c.707T>C; Exon 6) amino acid substitution mutation, the IVS8+1G>A (Exon 8) donor splice mutation, and the T416P (c.1246A>C; Exon 10) amino acid substitution mutation. These three mutations have been found in several cases of Pendred syndrome (Tsukamoto et al. 2003).
Pendred syndrome is an autosomal recessive disorder that accounts for 4-10% of the cases for hereditary deafness (Tesolin et al. 2021). Besides sensorineural hearing impairment, this syndrome is characterized by the presence of goiter, and a partial defect in iodide organification, which may be associated with insufficient thyroid hormone synthesis (Royaux et al. 2000). Goiter development (diffuse thyroid enlargement) and development of hypothyroidism are variable and depend on nutritional iodide intake. Pendred syndrome is caused by biallelic mutations in the SLC26A4
gene, which encodes pendrin, a transporter of chloride, bicarbonate and iodide. Mutations in this gene can also cause DFNB4 (Royaux et al. 2000). The SLC26A4
gene (21 exons) spans ~57.2 kb of genomic DNA and is located on chromosome 7q22.3, ~107 Mb from the p-telomere.
More information is available at https://www.ncbi.nlm.nih.gov/books/NBK1467/
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.
The SALSA MLPA Probemix P280-B4 SLC26A4 contains 38 MLPA probes with amplification products between 131 and 436 nucleotides (nt). This includes 24 probes for the SLC26A4
gene, with at least one probe for each exon. Among these, three probes are specific for the L236P (707T>C; Exon 6; 382 nt) mutation, the IVS8+1G>A (Exon 8; 321 nt) mutation, and the T416P (1246A>C; Exon 10; 227 nt) mutation, which will only generate a signal when the mutation is present. In addition, 14 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 (https://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 (see table below). More information on how to interpret observations on these control fragments can be found in the MLPA General Protocol and online at https://www.mrcholland.com
SALSA Binning DNA SD071
The SD071 Binning DNA provided with this probemix can be used for binning of all probes including the three mutation specific probes (227 nt probe 20624-L025811 for the T416P mutation, 321 nt probe 09249-L30464 for the IVS8+1G>A mutation, and 382 nt probe 09246-L13230 for the L236P mutation). SD071 Binning DNA is a mixture of genomic DNA from healthy individuals and plasmid DNA that contains the target sequence detected by the above mentioned probes. Inclusion of one reaction with 5 μl SD071 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 SD071 Binning DNA product description, available online: https://www.mrcholland.com
. This product is for research use only (RUO)
Sample DNA developed for this product: