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DNA sequence detection using surface-enhanced resonance Raman spectroscopy in a homogeneous multiplexed assay.
Anal Chem. 2009 Oct 01; 81(19):8134-40.AC

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.

Authors+Show Affiliations

Centre for Molecular Nanometrolgy, WestCHEM, Pure and Applied Chemistry, University of Strathclyde, 295 Cathedral Street, Glasgow, G1 1XL, UK.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19743872

Citation

MacAskill, Alexandra, et al. "DNA Sequence Detection Using Surface-enhanced Resonance Raman Spectroscopy in a Homogeneous Multiplexed Assay." Analytical Chemistry, vol. 81, no. 19, 2009, pp. 8134-40.
MacAskill A, Crawford D, Graham D, et al. DNA sequence detection using surface-enhanced resonance Raman spectroscopy in a homogeneous multiplexed assay. Anal Chem. 2009;81(19):8134-40.
MacAskill, A., Crawford, D., Graham, D., & Faulds, K. (2009). DNA sequence detection using surface-enhanced resonance Raman spectroscopy in a homogeneous multiplexed assay. Analytical Chemistry, 81(19), 8134-40. https://doi.org/10.1021/ac901361b
MacAskill A, et al. DNA Sequence Detection Using Surface-enhanced Resonance Raman Spectroscopy in a Homogeneous Multiplexed Assay. Anal Chem. 2009 Oct 1;81(19):8134-40. PubMed PMID: 19743872.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - DNA sequence detection using surface-enhanced resonance Raman spectroscopy in a homogeneous multiplexed assay. AU - MacAskill,Alexandra, AU - Crawford,David, AU - Graham,Duncan, AU - Faulds,Karen, PY - 2009/9/12/entrez PY - 2009/9/12/pubmed PY - 2009/12/18/medline SP - 8134 EP - 40 JF - Analytical chemistry JO - Anal Chem VL - 81 IS - 19 N2 - 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. SN - 1520-6882 UR - https://www.unboundmedicine.com/medline/citation/19743872/DNA_sequence_detection_using_surface_enhanced_resonance_Raman_spectroscopy_in_a_homogeneous_multiplexed_assay_ L2 - https://doi.org/10.1021/ac901361b DB - PRIME DP - Unbound Medicine ER -