Single-base extension and ELISA-based approach for single-nucleotide polymorphisms genotyping.
Single-nucleotide polymorphisms (SNPs) emerge as a fundamental tool in personalized medicine due to their association with drug responses or disease predisposition. Single-base extension (SBE) is a common method for characterizing known SNPs, but involves complicated procedures or requires costly analytical instruments. Here, we describe a novel SNP genotyping based on SBE and enzyme-linked immunosorbent assay (ELISA). During the SBE, the 5' end fluorescein isothiocyanate-labeled allele-specific primer will extend with biotinylated dideoxynucleotides which are complementary to the SNP sites. The extension product will then be captured by streptavidin-coated nanoparticle and develop blue color in the ELISA assay. We validated this method by detecting SNPs for TP53 gene codon 273 from 68 individuals and the data were 100% in concordant with DNA sequencing. Thus, SBE and ELISA-based SNPs assay is a simple and accurate method for SNP genotyping.
Liu
Guibo
G
Department of Anatomy, Mudanjiang Medical College, Mudanjiang, China.
Cheng
Yongxia
Y
Zhao
Wei
W
Jin
Zaishun
Z
Shan
Hongbo
H
Xu
Gaolian
G
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*