General information
The SALSA MLPA Probemix P465 ACADM is a research use only (RUO) assay for the detection of deletions or duplications in the ACADM gene, which is associated with Medium-Chain Acyl-Coenzyme A Dehydrogenase Deficiency (MCAD). This probemix can also be used to detect the presence of the wild-type sequence of c.985A>G/p.Lys329Glu (rs77931234) point mutation.

MCAD (OMIM 201450) is a disorder of fatty acid β-oxidation. Fatty acid β-oxidation fuels hepatic ketogenesis, a major source of energy for peripheral tissues once glycogen stores become depleted. This disorder is characterised by intolerance to prolonged fasting, impaired ketogenesis, low plasma and tissue carnitine levels, among others. The disorder may be severe, and even fatal, in young patients. Defects in the ACADM gene on chromosome 1 are the main cause of MCAD. Most of the known pathologic defects are point mutations but several articles describe small and large deletions of the ACADM gene.

More information is available at https://www.ncbi.nlm.nih.gov/books/NBK1424/.

Probemix content
The SALSA MLPA Probemix P465-A1 ACADM contains 27 MLPA probes with amplification products between 161 and 384 nucleotides (nt). This includes 15 probes for the ACADM gene, of which one probe specific for the wild-type sequence of the c.985A>G/p.Lys329Glu (rs77931234) mutation, which will only generate a signal when the wildtype sequence is present and three flanking probes that are located upstream and downstream of the ACADM gene. Furthermore, this probemix also contains nine reference probes 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.