DNA sequence detection using surface-enhanced resonance raman spectroscopy in a homogeneous multiplexed assay

A. Macaskill, D. Crawford, D. Graham, K. Faulds

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

Detection of specific DNA sequences is central to modern molecular biology and also to molecular diagnostics where identification of a particular disease is based on nucleic acid identification. Many methods exist, and fluorescence spectroscopy dominates the detection technologies employed with different assay formats. This study demonstrates the use of surface-enhanced resonance Raman scattering (SERRS) to detect specific DNA sequences when coupled with modified SERRS-active probes that have been designed to modify the affinity of double- and single-stranded DNA for the surface of silver nanoparticles resulting in discernible differences in the SERRS which can be correlated to the specific DNA hybridization event. The principle of the assay lies on the lack of affinity of double-stranded DNA for silver nanoparticle surfaces; therefore, hybridization of the probe to the target results in a reduction in the SERRS signal. Use of locked nucleic acid (LNA) residues in the DNA probes resulted in greater discrimination between exact match and mismatches when used in comparison to unmodified labeled DNA probes. Polymerase chain reaction (PCR) products were detected using this methodology, and ultimately a multiplex detection of sequences relating to a hospital-acquired infection, namely, methicillin-resistant Staphylococcus aureus (MRSA), demonstrated the versatility and applicability of this approach to real-life situations.
LanguageEnglish
Pages8134-8140
Number of pages7
JournalAnalytical Chemistry
Volume81
Issue number19
DOIs
Publication statusPublished - 10 Sep 2009

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DNA sequences
Raman spectroscopy
Assays
Raman scattering
DNA Probes
Silver
DNA
Nanoparticles
Molecular biology
Methicillin
Single-Stranded DNA
Polymerase chain reaction
Fluorescence spectroscopy
Reaction products
Nucleic Acids

Keywords

  • DNA
  • molecular diagnostics
  • silver nanoparticles
  • locked nucleic acid
  • polymerase chain reaction

Cite this

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title = "DNA sequence detection using surface-enhanced resonance raman spectroscopy in a homogeneous multiplexed assay",
abstract = "Detection of specific DNA sequences is central to modern molecular biology and also to molecular diagnostics where identification of a particular disease is based on nucleic acid identification. Many methods exist, and fluorescence spectroscopy dominates the detection technologies employed with different assay formats. This study demonstrates the use of surface-enhanced resonance Raman scattering (SERRS) to detect specific DNA sequences when coupled with modified SERRS-active probes that have been designed to modify the affinity of double- and single-stranded DNA for the surface of silver nanoparticles resulting in discernible differences in the SERRS which can be correlated to the specific DNA hybridization event. The principle of the assay lies on the lack of affinity of double-stranded DNA for silver nanoparticle surfaces; therefore, hybridization of the probe to the target results in a reduction in the SERRS signal. Use of locked nucleic acid (LNA) residues in the DNA probes resulted in greater discrimination between exact match and mismatches when used in comparison to unmodified labeled DNA probes. Polymerase chain reaction (PCR) products were detected using this methodology, and ultimately a multiplex detection of sequences relating to a hospital-acquired infection, namely, methicillin-resistant Staphylococcus aureus (MRSA), demonstrated the versatility and applicability of this approach to real-life situations.",
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DNA sequence detection using surface-enhanced resonance raman spectroscopy in a homogeneous multiplexed assay. / Macaskill, A.; Crawford, D.; Graham, D.; Faulds, K.

In: Analytical Chemistry, Vol. 81, No. 19, 10.09.2009, p. 8134-8140.

Research output: Contribution to journalArticle

TY - JOUR

T1 - DNA sequence detection using surface-enhanced resonance raman spectroscopy in a homogeneous multiplexed assay

AU - Macaskill, A.

AU - Crawford, D.

AU - Graham, D.

AU - Faulds, K.

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Y1 - 2009/9/10

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AB - Detection of specific DNA sequences is central to modern molecular biology and also to molecular diagnostics where identification of a particular disease is based on nucleic acid identification. Many methods exist, and fluorescence spectroscopy dominates the detection technologies employed with different assay formats. This study demonstrates the use of surface-enhanced resonance Raman scattering (SERRS) to detect specific DNA sequences when coupled with modified SERRS-active probes that have been designed to modify the affinity of double- and single-stranded DNA for the surface of silver nanoparticles resulting in discernible differences in the SERRS which can be correlated to the specific DNA hybridization event. The principle of the assay lies on the lack of affinity of double-stranded DNA for silver nanoparticle surfaces; therefore, hybridization of the probe to the target results in a reduction in the SERRS signal. Use of locked nucleic acid (LNA) residues in the DNA probes resulted in greater discrimination between exact match and mismatches when used in comparison to unmodified labeled DNA probes. Polymerase chain reaction (PCR) products were detected using this methodology, and ultimately a multiplex detection of sequences relating to a hospital-acquired infection, namely, methicillin-resistant Staphylococcus aureus (MRSA), demonstrated the versatility and applicability of this approach to real-life situations.

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KW - molecular diagnostics

KW - silver nanoparticles

KW - locked nucleic acid

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