General information: The SALSA MLPA
Probemix P169 Hirschsprung-1 is a
research use only (RUO) assay for the detection of deletions or duplications in the
RET,
ZEB2,
EDN3 and
GDNF genes, which are associated with Hirschsprung disease (HSCR). HSCR or aganglionic megacolon, is a congenital disorder characterised by the absence of enteric ganglia along a variable length of the intestine. The disease can be classified according to the length of the aganglionosis into short segment (accounts for 80% of the HSCR cases), long segment and total colonic aganglionosis forms. Dominant mutations in the
RET gene, as well as recessive mutations in several other genes including the
ZEB2,
EDN3, and
GDNF genes, are associated with HSCR.
The
RET gene encodes for one of the cell-surface receptor tyrosine kinases, which plays a role in cell growth and differentiation.
RET is composed of 20 exons, spans ~53 kb of genomic DNA, and is located on 10q11.21, ~43 Mb from the p-telomere. The
ZEB2 gene encodes for a member of the Zfh1 family and functions as a DNA-binding transcriptional repressor that interacts with activated SMADs (SMADs are the main signal transducers for receptors of the TGF-β superfamily). The
EDN3 gene encodes for a member of the endothelin family.
EDN3 consists of 5 exons, spans ~26 kb of genomic DNA, and is located on 20q13.32, ~59 Mb from the p-telomere. The
GDNF gene encodes for a neurotrophic factor.
GDNF consists of 3 exons, spans ~27 kb of genomic DNA and is located on 5p13.2, ~38 Mb from the p-telomere.
More information on HSCR is available at
https://www.ncbi.nlm.nih.gov/books/NBK1439/.
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 P169-C2 Hirschsprung-1 contains 51 MLPA probes with amplification products between 130 and 490 nucleotides (nt). This includes 12 probes for the
ZEB2 gene, five probes for
GDNF, 20 probes for
RET, and five probes for
EDN3. In addition, nine 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.mlpa.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.mlpa.com.