Abstract
| Language | English |
|---|---|
| Pages | 8134-8140 |
| Number of pages | 7 |
| Journal | Analytical Chemistry |
| Volume | 81 |
| Issue number | 19 |
| DOIs | |
| Publication status | Published - 10 Sep 2009 |
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Keywords
- DNA
- molecular diagnostics
- silver nanoparticles
- locked nucleic acid
- polymerase chain reaction
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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 journal › Article
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.
PY - 2009/9/10
Y1 - 2009/9/10
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.
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.
KW - DNA
KW - molecular diagnostics
KW - silver nanoparticles
KW - locked nucleic acid
KW - polymerase chain reaction
U2 - 10.1021/ac901361b
DO - 10.1021/ac901361b
M3 - Article
VL - 81
SP - 8134
EP - 8140
JO - Analytical Chemistry
T2 - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 19
ER -