Journal Article

Assay of Single Nucleotide Polymorphisms Based on Ligase Detection Reaction and Enzyme-Linked Immunosorbent Assay

Xiaodong Zhang, Jianbo Zhang, Xiumei Yuan, Jialiang Wang, Changhao Yin, Decai Fu, Chunyan Yu, Dawei Zhang, Hengjuan Lu and Jingchao Li

in Laboratory Medicine

Published on behalf of American Society for Clinical Pathology

Volume 44, issue 2, pages e10-e12
Published in print May 2013 | ISSN: 0007-5027
Published online October 2015 | e-ISSN: 1943-7730 | DOI: https://dx.doi.org/10.1309/LMM4LPLC53KIRKMV
Assay of Single Nucleotide Polymorphisms Based on Ligase Detection Reaction and Enzyme-Linked Immunosorbent Assay

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  • Clinical Cytogenetics and Molecular Genetics
  • Chemistry
  • Molecular and Cell Biology
  • Molecular Biology and Genetics

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Ligase detection reaction (LDR)–based methods are usually expensive procedures for detecting known single nucleotide polymorphisms (SNPs). Herein, we describe a novel and cost-effective genotyping method for detection of SNPs based on LDR and enzyme-linked immunosorbent assay (ELISA). PCR is used to amplify the region containing the site of the SNPs. In LDR, the 5′ end biotin-labeled allele specific discrimination probe and the 3′ end fluorescein isothiocyanate-labeled detector probe will, when mediated by a thermal DNA ligase, link together. The LDR product will then be separated using streptavidin-coated nanoparticles and develop a blue color during ELISA. We used the LDR-ELISA method to detect SNPs for p53 gene codon 282 in 50 individuals. Our results showed 100% concordance between LDR-ELISA and DNA sequencing. Therefore, LDR-ELISA SNPs assay is a simple, accurate tool for SNP genotyping.

Keywords: single nucleotide polymorphisms; ligase detection reaction; ELISA

Journal Article.  1305 words.  Illustrated.

Subjects: Clinical Cytogenetics and Molecular Genetics ; Chemistry ; Molecular and Cell Biology ; Molecular Biology and Genetics

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