Multiplexed detection of six labelled oligonucleotides using surface enhanced resonance Raman scattering (SERRS)

K. Faulds, R. Jarvis, W.E. Smith, D. Graham, R. Goodacre

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

The labelling of target biomolecules followed by detection using some form of optical spectroscopy has become common practice to aid in their detection. This approach has allowed the field of bioanalysis to dramatically expand; however, most methods suffer from the lack of the ability to discriminate between the components of a complex mixture. Currently, fluorescence spectroscopy is the method of choice but its ability to multiplex is greatly hampered by the broad overlapping spectra which are obtained. Surface enhanced resonance Raman scattering (SERRS) holds many advantages over fluorescence both in sensitivity and, more importantly here, in its ability to identify components in a mixture without separation due to the sharp fingerprint spectra obtained. Here the first multiplexed simultaneous detection of six different DNA sequences, corresponding to different strains of the Escherichia coli bacterium, each labelled with a different commercially available dye label (ROX, HEX, FAM, TET, Cy3, or TAMRA) is reported. This was achieved with the aid of multivariate analysis, also known as chemometrics, which can involve the application of a wide range of statistical and data analysis methods. In this study, both exploratory discriminant analysis and supervised learning, by partial least squares (PLS) regression, were used and the ability to discriminate whether a particular labelled oligonucleotide was present or absent in a mixture was achieved using PLS with very high sensitivity (0.98-1), specificity (0.98-1), accuracy (range 0.99-1), and precision (0.98-1). (Abstract copied from Swetswise web site: http://www.swetswise.com/eAccess/viewAbstract.do?articleID=38598265)
LanguageEnglish
Pages1505-1512
Number of pages8
JournalAnalyst
Volume133
Issue number11
Early online date28 Aug 2008
DOIs
Publication statusPublished - 1 Nov 2008

Fingerprint

Aptitude
Raman Spectrum Analysis
Oligonucleotides
Raman scattering
scattering
DNA sequences
Supervised learning
Fluorescence spectroscopy
Biomolecules
Discriminant analysis
Least-Squares Analysis
Complex Mixtures
Labeling
Escherichia coli
Labels
Statistical methods
Bacteria
Coloring Agents
fluorescence spectroscopy
Fluorescence

Keywords

  • multiplexed detection
  • labelled oligonucleotides
  • surfaceenhanced resonance
  • Raman scattering (SERRS)†

Cite this

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Multiplexed detection of six labelled oligonucleotides using surface enhanced resonance Raman scattering (SERRS). / Faulds, K.; Jarvis, R.; Smith, W.E.; Graham, D.; Goodacre, R.

In: Analyst, Vol. 133, No. 11, 01.11.2008, p. 1505-1512.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Multiplexed detection of six labelled oligonucleotides using surface enhanced resonance Raman scattering (SERRS)

AU - Faulds, K.

AU - Jarvis, R.

AU - Smith, W.E.

AU - Graham, D.

AU - Goodacre, R.

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AB - The labelling of target biomolecules followed by detection using some form of optical spectroscopy has become common practice to aid in their detection. This approach has allowed the field of bioanalysis to dramatically expand; however, most methods suffer from the lack of the ability to discriminate between the components of a complex mixture. Currently, fluorescence spectroscopy is the method of choice but its ability to multiplex is greatly hampered by the broad overlapping spectra which are obtained. Surface enhanced resonance Raman scattering (SERRS) holds many advantages over fluorescence both in sensitivity and, more importantly here, in its ability to identify components in a mixture without separation due to the sharp fingerprint spectra obtained. Here the first multiplexed simultaneous detection of six different DNA sequences, corresponding to different strains of the Escherichia coli bacterium, each labelled with a different commercially available dye label (ROX, HEX, FAM, TET, Cy3, or TAMRA) is reported. This was achieved with the aid of multivariate analysis, also known as chemometrics, which can involve the application of a wide range of statistical and data analysis methods. In this study, both exploratory discriminant analysis and supervised learning, by partial least squares (PLS) regression, were used and the ability to discriminate whether a particular labelled oligonucleotide was present or absent in a mixture was achieved using PLS with very high sensitivity (0.98-1), specificity (0.98-1), accuracy (range 0.99-1), and precision (0.98-1). (Abstract copied from Swetswise web site: http://www.swetswise.com/eAccess/viewAbstract.do?articleID=38598265)

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