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Visual Detection of Multiple Nucleic Acids in a Capillary Array
Published on: November 15, 2017
Multiplex single nucleotide polymorphism genotyping utilizing ligase detection reaction coupled surface enhanced
Adam J Lowe1, Yun Suk Huh, Aaron D Strickland
1Graduate Field of Microbiology, Cornell University, Ithaca, New York 14853, USA. ajl248@cornell.edu
Analytical Chemistry
|June 10, 2010
Summary
This study introduces a novel multiplex method combining ligase detection reaction (LDR) and surface-enhanced Raman spectroscopy (SERS) for accurate single nucleotide polymorphism (SNP) genotyping. The platform demonstrates high sensitivity and quantitative correlation for K-Ras oncogene allele detection.
Area of Science:
- Biotechnology
- Genomics
- Spectroscopy
Background:
- Single nucleotide polymorphisms (SNPs) are crucial biomarkers for genetic diseases, cancer, and pharmacogenomics.
- Ligase detection reaction (LDR) offers high specificity and low detection limits for SNP identification.
- Multiplex SNP genotyping is essential for comprehensive genetic analysis.
Purpose of the Study:
- To develop and validate the first multiplex ligase detection reaction-surface enhanced Raman spectroscopy (LDR-SERS) SNP genotyping scheme.
- To optimize DNA labeling and Raman spectroscopy conditions for enhanced sensitivity and distinct diagnostic peaks.
- To demonstrate the quantitative correlation between diagnostic Raman peak intensity and SNP target concentration in heterozygous samples.
Main Methods:
- Development of a multiplex LDR-SERS platform for simultaneous SNP detection.
- Optimization of DNA labeling strategies and Raman spectroscopy parameters.
- Genotyping of K-Ras oncogene alleles using genomic DNA from cell lines at 10 pM detection levels.
Main Results:
- Achieved 10 pM detection levels for K-Ras oncogene allele genotyping.
- Demonstrated distinct diagnostic Raman peaks, offering advantages over fluorescence detection.
- Established a linear correlation between diagnostic peak intensity and SNP target concentration in heterozygous samples.
Conclusions:
- The developed LDR-SERS platform enables sensitive and multiplexed SNP genotyping.
- Raman spectroscopy provides distinct and quantifiable signals for genetic analysis.
- This technology advances SNP genotyping capabilities for applications in genetic disease and cancer research.
