Simple multiplex genotyping by surface-enhanced resonance Raman scattering

Analytical Chemistry
Duncan GrahamW Ewen Smith

Abstract

The accurate detection of DNA sequences is essential for a variety of post human genome projects including detection of specific gene variants for medical diagnostics and pharmacogenomics. A specific DNA sequence detection assay based on surface-enhanced resonance Raman scattering (SERRS) and an amplification refractory mutation system (ARMS) is reported. Initially, generation of PCR products was achieved by using specifically designed allele-specific SERRS active primers. Detection by SERRS of the PCR products confirmed the presence of the sequence tested for by the allele-specific oligonucleotides. This lead directly to the multiplex genotyping of human DNA samples for the deltaF508 mutational status of the cystic fibrosis transmembrane conductance regulator gene using SERRS active primers in an ARMS assay. Removal of the unincorporated primers allowed fast and accurate analysis of the three genotypes possible in this system in a multiplex format without any separation of amplicons. The results indicate that SERRS can be used in modern genetic analysis and offers an opportunity for the development of novel assays. This is the first demonstration of the use of SERRS in multiplex genotyping and shows potential advantages over f...Continue Reading

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Citations

May 16, 2009·Analytical and Bioanalytical Chemistry·Guoqing WangAndrew J deMello
Jun 23, 2009·Analytical and Bioanalytical Chemistry·Magdalena GellnerSebastian Schlücker
Jun 15, 2012·Analytical and Bioanalytical Chemistry·Cécile FeuillieIsabelle Daniel
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Jan 24, 2020·The Analyst·Daniel MacdonaldDuncan Graham

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