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. 2007 Jun 1;79(11):3981-8.
doi: 10.1021/ac070078z. Epub 2007 Apr 28.

Surface-enhanced Raman scattering based nonfluorescent probe for multiplex DNA detection

Lan Sun  1 , Chenxu Yu, Joseph Irudayaraj
Affiliations

Surface-enhanced Raman scattering based nonfluorescent probe for multiplex DNA detection

Lan Sun et al. Anal Chem. .

Abstract

To provide rapid and accurate detection of DNA markers in a straightforward, inexpensive, and multiplex format, an alternative surface-enhanced Raman scattering based probe was designed and fabricated to covalently attach both DNA probing sequence and nonfluorescent Raman tags to the surface of gold nanoparticles (DNA-AuP-RTag). The intensity of Raman signal of the probes could be controlled through the surface coverage of the nonfluorescent Raman tags (RTags). Detection sensitivity of these probes could be optimized by fine-tuning the amount of DNA molecules and RTags on the probes. Long-term stability of the DNA-AuP-RTag probes was found to be good (over 3 months). Excellent multiplexing capability of the DNA-AuP-RTag scheme was demonstrated by simultaneous identification of up to eight probes in a mixture. Detection of hybridization of single-stranded DNA to its complementary targets was successfully accomplished with a long-term goal to use nonfluorescent RTags in a Raman-based DNA microarray platform.

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Figures

Figure 1
Figure 1
Fabrication of the DNA-AuP-RTag probe.
Figure 2
Figure 2
Structures of selected non-fluorescent Raman tags (RTags).
Figure 3
Figure 3
Raman spectra of gold nanoparticles functionalized with 1 μM DNA (control has no DNA).
Figure 4
Figure 4
Raman spectra of DNA-AuP-RTag probes.
Figure 5
Figure 5
Variation in Raman intensity of the peak at 1089 cm−1 w/r/t 4-mercaptopyridine concentration.
Figure 6
Figure 6
Multiplex detection of DNA-AuP-RTag probe mixtures. Mixture-1: DNA-AuP-RTag-1 & 2; Mixture-2: DNA-AuP-RTag-1, 2, 3 & 4; Mixture-3: DNA-AuP-RTag-1 to 8.
Figure 7
Figure 7
Change in absorbance in the visible region of the UV-visible spectra for (A) mixture of DNA1 (5'-thiol-AAA AAA AAA GCA GCC AAT TC-3', eight A spacers) and DNA2 (5'-thiol-AAA AAA AAG AAT TGG CTG CT-3' containing eight A spacers followed by the complementary strand) functionalized gold nanoparticles (B) mixture of DNA1-AuP-RTag-1 and DNA2 functionalized gold nanoparticles (C) gold nanoparticles functionalized with ssDNA molecules at different temperature (D) the DNA1-AuP-RTag-1 probe at different temperature.
Figure 8
Figure 8
Long-term stability of the DNA-AuP-RTag probes.
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