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Multiplex Single Nucleotide Polymorphism Genotyping Utilizing Ligase Detection Reaction Coupled Surface Enhanced Raman Spectroscopy

NCJ Number
308445
Journal
Analytical Chemistry Volume: 82 Issue: 13 Dated: 2010 Pages: 5810–5814
Author(s)
Adam J. Lowe; Yun Suk Huh; Aaron D. Strickland; David Erickson; Carl A. Batt
Date Published
2010
Length
5 pages
Annotation

This study explores the use of multiplex single nucleotide polymorphism genotyping through a ligase detection reaction coupled surface enhanced raman spectroscopy.

Abstract

This paper presents the first multiplex LDR (ligase detection reaction)-surface enhanced Raman spectroscopy (SERS) SNP genotyping scheme. The platform has the advantage in that the diagnostic peaks of Raman are more distinct than fluorescence, and in theory, a clinically significant number of markers can be multiplexed in a single sample using different SERS reporters. Here researchers report LDR-SERS multiplex SNP genotyping of K-Ras oncogene alleles at 10 pM detection levels, optimization of DNA labeling as well as Raman conditions, and the linear correlation of diagnostic peak intensity to SNP target concentration in heterozygous samples. Genomic DNA from typed cells lines was obtained and scored for the K-Ras genotype. These advances are significant, as the researchers have further developed their new SNP genotyping platform and have demonstrated the ability to correlate genotype ratios directly to diagnostic Raman peak signal intensity. Single nucleotide polymorphisms (SNPs) are one of the key diagnostic markers for genetic disease, cancer progression, and pharmacogenomics. The (LDR) is an excellent method to identify SNPs, combining low detection limits and high specificity. (Published Abstract Provided)