The SALSA MLPA Probemix P165 HSP mix-1 is an in vitro diagnostic (IVD) or research use only (RUO) semi-quantitative manual assay for the detection of deletions and duplications in SPAST gene, in order to confirm a potential cause for and clinical diagnosis of spastic paraplegia (SPG) type 4, and for molecular genetic testing of at-risk family members. This assay is for use with genomic DNA isolated from human peripheral whole blood specimens.

Certain probes targeting additional genes included in P165 HSP mix-1 may only be used in a research setting. The following table summarises which probes are for IVD, and which are exclusively restricted to RUO use.

For the full intended purpose, see the product description.

Hereditary spastic paraplegias (HSP) are genetically heterogeneous neurodegenerative disorders characterised by progressive spasticity and weakness of the lower limbs due to axonal degeneration in the pyramidal tract. To date, more than 80 genetic types of HSP have been defined by genetic linkage analysis and identification of HSP-related gene variants.

Mutations in the SPAST gene are responsible for both autosomal dominant HSP (40–50%) and sporadic cases (10–15%), causing spastic paraplegia type 4 (SPG4). The most common type of SPAST mutations are point mutations (75-80%), and large genomic abnormalities, such as exon deletions, account for up to 20-25% of disease-associated SPAST mutations (Beetz et al. 2006; d'Amore et al. 2018; Depienne et al. 2006; Kadnikova et al. 2019).

SALSA MLPA Probemix P165 HSP mix-1 is CE-marked under the IVDR for in vitro diagnostic (IVD) use in Europe.

This assay is for research use only (RUO) in all other territories.

Inclusion of a positive sample is usually not required, but can be useful for the analysis of your experiments. MRC Holland has very limited access to positive samples and cannot supply such samples. We recommend using positive samples from your own collection. Alternatively, you can use positive samples from an online biorepository, such as the Coriell Institute.

The commercially available positive samples below have been tested with the current (C3) version of this product and have been shown to produce useful results.

• Coriell HG00500: Heterozygous duplication affecting probes for SPAST exon 4-17.
• Coriell NA10401: Heterozygous duplication affecting all SPAST probes.

Key related products

• P213 HSP mix-2: Targets REEP1, and SPG7, involved in spastic paraplegia type 7 (SPG7) and type 31 (SPG31).
• P211 HSP region: Targets SPAST, the 2p22.3 region, and the 15q11.2 region, involved in hereditary spastic paraplegias (HSPs).
• P306 SPG11: Targets SPG11, involved in spastic paralegia type 11 (SPG11).

General MLPA resources

• MLPA technique overview
• Coffalyser.Net overview

MLPA education

• Getting started with MLPA
• Using Coffalyser.Net to analyse MLPA data
• Help centre with other learning resources

Selected publications using P165 HSP mix-1

• D'Amore A et al. (2018). Next Generation Molecular Diagnosis of Hereditary Spastic Paraplegias: An Italian Cross-Sectional Study. Front Neurol. 9:981.
• Hashemi SS et al. (2022). Anticipation Can Be More Common in Hereditary Spastic Paraplegia with SPAST Mutations Than It Appears. Can J Neurol Sci. 49:651-61.
• Kadnikova VA et al. (2019). Mutational Spectrum of Spast (Spg4) and Atl1 (Spg3a) Genes In Russian Patients With Hereditary Spastic Paraplegia. Sci Rep. 9:14412.
• Magariello A et al. (2010). Mutation analysis of the SPG4 gene in Italian patients with pure and complicated forms of spastic paraplegia. J Neurol Sci. 288:96-100.
• Mészárosová AU et al. (2016). SPAST mutation spectrum and familial occurrence among Czech patients with pure hereditary spastic paraplegia. J Hum Genet. 61:845-50.
• Navas-Sánchez FJ et al. (2021). Thalamic atrophy in patients with pure hereditary spastic paraplegia type 4. J Neurol. 268:2429-40.
• Park H et al. (2015). Mutational spectrum of the SPAST and ATL1 genes in Korean patients with hereditary spastic paraplegia. J Neurol Sci. 357:167-72.
• Parodi L et al. (2018). Spastic paraplegia due to SPAST mutations is modified by the underlying mutation and sex. Brain. 141:3331-42.
• Sulek A et al. (2013). Screening for the hereditary spastic paraplaegias SPG4 and SPG3A with the multiplex ligation-dependent probe amplification technique in a large population of affected individuals. Neurol Sci. 34:239-42.

References

• Beetz C et al. (2006). High frequency of partial SPAST deletions in autosomal dominant hereditary spastic paraplegia. Neurology. 67:1926-30.
• D'Amore A et al. (2018). Next Generation Molecular Diagnosis of Hereditary Spastic Paraplegias: An Italian Cross-Sectional Study. Front Neurol. 9:981.
• Depienne C et al. (2007). Exon deletions of SPG4 are a frequent cause of hereditary spastic paraplegia. J Med Genet. 44:281-4.
• Kadnikova VA et al. (2019). Mutational Spectrum of Spast (Spg4) and Atl1 (Spg3a) Genes In Russian Patients With Hereditary Spastic Paraplegia. Sci Rep. 9:14412.
• Sulek A et al. (2013). Screening for the hereditary spastic paraplaegias SPG4 and SPG3A with the multiplex ligation-dependent probe amplification technique in a large population of affected individuals. Neurol Sci. 34:239-42.