The SALSA MLPA Probemix P016 VHL is an in vitro diagnostic (IVD) or research use only (RUO) semi-quantitative assay for the detection of deletions in the VHL gene in genomic DNA isolated from human peripheral whole blood specimens. P016 VHL is intended to confirm a potential cause for and clinical diagnosis of Von Hippel-Lindau (VHL) disease and for molecular genetic testing of at-risk family members.

For the full intended purpose, see the product description.

Von Hippel-Lindau (VHL) disease is a dominantly inherited familial cancer syndrome predisposing to a variety of malignant and benign neoplasms, most frequently retinal, cerebellar and spinal hemangioblastoma, renal cell carcinoma, pheochromocytoma, and pancreatic tumours. The basis of familial inheritance of VHL disease is a germline mutation in the VHL tumour suppressor gene, located in chromosomal region 3p25.3. Approximately 80% of individuals with VHL disease inherit a mutation, whereas 20% of individuals have a de novo mutation (http://www.ncbi.nlm.nih.gov/books/NBK1463/). Of the mutations in the VHL gene, 30-60% are missense mutations, 20–40% are large intragenic deletions (0.5–250 kb), 12–20% are microdeletions or insertions and 7–11% are nonsense mutations (Decker et al. 2014).

Interestingly, loss of the nearby BRK1 gene in combination with loss of (part of) the VHL gene can be associated with a reduced risk of renal cell carcinoma as compared to defects in the VHL gene only (Escobar et al. 2010, McNeill et al. 2009). This probemix contains two probes that target the BRK1 gene.

SALSA MLPA Probemix P016 VHL is CE-marked under the IVDD for in vitro diagnostic (IVD) use in Europe. This assay has also been registered for IVD use in Israel.

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 (C2) version of this product and have been shown to produce useful results.

• Coriell NA10985: Heterozygous deletion affecting the probes for CNTN6, FANCD2, BRK1, VHL, IRAK2 and GHRL.
• Coriell NA13249: Heterozygous deletion affecting the probes for VHL exon 2-3 and IRAK2.
• Coriell NA13250: Heterozygous deletion affecting the probes for VHL exon 1-2.
• Coriell NA13256: Heterozygous deletion affecting the probes for FANCD2, BRK1 and VHL.

Key related products

• P226 SDH: Targets SDHB, SDHC, SDHD, SDHAF1, and SDHAF2, involved in hereditary paraganglioma/pheochromocytoma (PGL/PCC).
• P429 SDHA-MAX-TMEM127: Targets SDHA, MAX, and TMEM127, involved in pheochromocytomas and paragangliomas.

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 P016 VHL

• Franke G et al. (2009). Alu-Alu recombination underlies the vast majority of large VHL germline deletions: Molecular characterization and genotype-phenotype correlations in VHL patients. Hum Mutat. 30:776-86.
• Hes FJ et al. (2007). Frequency of Von Hippel-Lindau germline mutations in classic and non-classic Von Hippel-Lindau disease identified by DNA sequencing, Southern blot analysis and multiplex ligation-dependent probe amplification. Clin Genet. 72:122-9.
• Huang JS et al. (2007). Associations between VHL genotype and clinical phenotype in familial von Hippel-Lindau disease. Eur J Clin Invest. 37:492-500.
• McNeill A et al. (2009). Genotype-phenotype correlations in VHL exon deletions. Am J Med Genet A. 149A:2147-51.
• Tamura K et al. (2023). Variant spectrum of von Hippel-Lindau disease and its genomic heterogeneity in Japan. Hum Mol Genet. 32:2046-54.
• Vikkath N et al. (2015). Genotype-phenotype analysis of von Hippel-Lindau syndrome in fifteen Indian families. Fam Cancer. 14:585-94.
• Zhang K et al. (2023). Genotype-phenotype correlations and clinical outcomes of patients with von Hippel-Lindau disease with large deletions. J Med Genet. 60:477-83.

References

• Decker J et al. (2014). Clinical utility gene card for: von Hippel-Lindau (VHL). Eur J Hum Genet. 22.
• Escobar B et al. (2010). Brick1 is an essential regulator of actin cytoskeleton required for embryonic development and cell transformation. Cancer Res. 70:9349-59.
• McNeill A et al. (2009). Genotype-phenotype correlations in VHL exon deletions. Am J Med Genet A. 149A:2147-51.