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Analytical Validation of SOD2 Genotyping.
Marija Debeljak1, Stacy Riel1, Ming-Tseh Lin1
1Department of Pathology, The Sol Goldman Pancreatic Cancer Research Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
Methods and Protocols
|January 17, 2023
Summary
A new pyrosequencing assay accurately identifies a common genetic variation in the Manganese superoxide dismutase-2 (SOD2) gene. This assay helps identify prostate cancer patients eligible for antioxidant therapy clinical trials.
Area of Science:
- Genetics
- Biochemistry
- Oncology
Background:
- Manganese superoxide dismutase-2 (SOD2) protects cells from mitochondrial oxidative damage.
- A SOD2 gene polymorphism is linked to diseases, including prostate cancer.
- Specific SOD2 genotypes may influence treatment response to antioxidants in prostate cancer patients.
Purpose of the Study:
- To develop and validate a pyrosequencing assay for the common SOD2 single nucleotide polymorphism (SNP) rs4880.
- To identify men with castrate-resistant prostate cancer eligible for an antioxidant therapy clinical trial.
Main Methods:
- A pyrosequencing assay was developed to detect the SOD2 rs4880 SNP.
- 37 samples with known genotypes (confirmed by Illumina and Sanger sequencing) were used for validation.
- The assay was performed in a blinded manner on the selected samples.
Main Results:
- The pyrosequencing assay demonstrated 100% concordance with established genotyping methods for all 37 samples.
- Genotypes included homozygous Ala, heterozygous Ala/Val, and homozygous Val.
- The assay has been operational since May 2018, proving robust and accurate.
Conclusions:
- A validated pyrosequencing assay for SOD2 rs4880 is available.
- This assay can facilitate patient selection for clinical trials involving antioxidant therapies.
- The assay supports precision medicine approaches in prostate cancer treatment.
