Toward PCR-free mutation detection based on surface-enhanced Raman scattering

Mustafa Culha; Omer F. Karatas; Omer Aydin; Mehmet Kahraman; Kemal Keseroğlu; Ismail Sayin; Omer F. Bayrak

doi:10.1117/12.808267

17 February 2009 Toward PCR-free mutation detection based on surface-enhanced Raman scattering

Mustafa Culha, Omer F. Karatas, Omer Aydin, Mehmet Kahraman, Kemal Keseroğlu, Ismail Sayin, Omer F. Bayrak

Proceedings Volume 7192, Plasmonics in Biology and Medicine VI; 719205 (2009) https://doi.org/10.1117/12.808267
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

The development of an assay for the detection of gene mutations has been attempted based on surface-enhanced Raman scattering (SERS). Using multiplexing property and high sensitivity of SERS technique, the detection of all mutation possibilities on one given spot is achievable. To test the feasibility of approach, SNPs and other types of mutations such as insertion and deletion are investigated. The PCR amplified and isolated genomic DNA without PCR amplification is immobilized on poly-L/D-lysine coated glass surface after denaturing with heating. The SERS probes are prepared by simultaneous attachment of oligonucleotides complementary to the target mutation regions and Raman active dyes to 13 nm gold nanoparticles (GNPs). After the hybridization of SERS probes on the poly-L/D-lysine surfaces, it was stained with silver colloidal nanoparticles for further enhancement of Raman scattering. In the second approach, Raman active dyes are chemically attached on gold nanoparticles and a thin layer of silver film is deposited on top of it to prepare core-shell nanoparticles. The complementary oligonucleotides to the target regions of the gene are chemically attached to silver surfaces of the nanoparticles. The promising results indicate that it is possible to detect certain mutation types without PCR amplification using the approach.

Citation Download Citation

Mustafa Culha, Omer F. Karatas, Omer Aydin, Mehmet Kahraman, Kemal Keseroğlu, Ismail Sayin, and Omer F. Bayrak "Toward PCR-free mutation detection based on surface-enhanced Raman scattering", Proc. SPIE 7192, Plasmonics in Biology and Medicine VI, 719205 (17 February 2009); https://doi.org/10.1117/12.808267

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KEYWORDS

Nanoparticles

Gold

Raman spectroscopy

Silver

Raman scattering

Molecules

Rhodamine

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