Single-Base Extension and ELISA-Based Approach for Single-Nucleotide Polymorphisms Genotyping
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- 作者：Guibo Liu (1)
Yongxia Cheng (2)
Wei Zhao (1)
Zaishun Jin (2) zaishun5@yahoo.com
Hongbo Shan (3)
Gaolian Xu (4) - 关键词：Single ; nucleotide polymorphisms – ; Genotyping – ; ELISA – ; Single ; base extension
- 刊名：Applied Biochemistry and Biotechnology
- 出版时间：March 2011
- 出版年：2011
- 期刊代码：pc312_0273-2289
- 类别：ch
- 卷：163
- 期：5
- 页码：573-576
- 数据来源：sp
摘要
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.