Analytical Validation of <i>SOD2</i> Genotyping

Manganese superoxide dismutase-2 (<i>SOD2</i>) plays a crucial role in cells’ protection against mitochondrial oxidative damage. A genetic polymorphism in the mitochondrial targeting sequence of the <i>SOD2</i> gene has been implicated in various diseases, including prostate...

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Bibliographic Details
Main Authors: Marija Debeljak, Stacy Riel, Ming-Tseh Lin, James R. Eshleman, Channing J. Paller
Format: Article
Language:English
Published: MDPI AG 2022-12-01
Series:Methods and Protocols
Subjects:
Online Access:https://www.mdpi.com/2409-9279/6/1/4
Description
Summary:Manganese superoxide dismutase-2 (<i>SOD2</i>) plays a crucial role in cells’ protection against mitochondrial oxidative damage. A genetic polymorphism in the mitochondrial targeting sequence of the <i>SOD2</i> gene has been implicated in various diseases, including prostate cancer. Paller et al. have shown an increase in prostate-specific antigen (PSA) doubling time in patients with the Ala/Ala (wildtype) genotype when treated with pomegranate/grape extract antioxidants. We developed and validated a pyrosequencing assay that detects the common germline <i>SOD2</i> SNP (rs_4880) with the aim of identifying men with castrate-resistant prostate cancer eligible for an antioxidant therapy clinical trial. We first selected 37 samples from the 1000 genomes study with known genotypes determined using Illumina-based sequencing and confirmed them by Sanger sequencing. In a blinded design, we then performed the new pyrosequencing assay on these samples and assigned genotypes. Genotypes for all 37 samples (13 homozygous Ala, 12 heterozygous Ala/Val, and 12 homozygous Val) were all concordant by pyrosequencing. The pyrosequencing assay has been live since May 2018 and has proven to be robust and accurate.
ISSN:2409-9279