Simple multiplex genotyping by surface-enhanced resonance Raman scattering

D. Graham, B.J. Mallinder, D. Whitcombe, N.D. Watson, W.E. Smith

Research output: Contribution to journalArticle

135 Citations (Scopus)

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 DeltaF(508) 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 fluorescence as a detection technique,with considerable promise for future development.
LanguageEnglish
Pages1069-1074
Number of pages5
JournalAnalytical Chemistry
Volume74
Issue number5
DOIs
Publication statusPublished - 1 Mar 2002

Fingerprint

Raman scattering
Assays
Genes
DNA sequences
Refractory materials
Amplification
Cystic Fibrosis Transmembrane Conductance Regulator
Oligonucleotides
Demonstrations
Fluorescence
DNA

Keywords

  • colloidal silver particles
  • labeled DNA
  • fluorescence detection
  • nucleic-acids
  • spectroscopy
  • sers
  • adsorption
  • dyes

Cite this

Graham, D. ; Mallinder, B.J. ; Whitcombe, D. ; Watson, N.D. ; Smith, W.E. / Simple multiplex genotyping by surface-enhanced resonance Raman scattering. In: Analytical Chemistry. 2002 ; Vol. 74, No. 5. pp. 1069-1074.
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Simple multiplex genotyping by surface-enhanced resonance Raman scattering. / Graham, D.; Mallinder, B.J.; Whitcombe, D.; Watson, N.D.; Smith, W.E.

In: Analytical Chemistry, Vol. 74, No. 5, 01.03.2002, p. 1069-1074.

Research output: Contribution to journalArticle

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