期刊
BIOSENSORS & BIOELECTRONICS
卷 31, 期 1, 页码 451-457出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2011.11.014
关键词
Methylation; Single base extension; Surface enhanced Raman spectroscopy; DNA; Sensitivity
类别
资金
- Chinese Academy of Sciences [KGCX2-YW-130]
- National Natural Science Foundation of China [21075129]
- National Basic Research Program 973 [2011CB933600, 2010CB732600]
DNA methylation is a key diagnostic maker for genetic disease, cancer progression and pharmcogenomics. So far various techniques have been developed for DNA methylation assay, but most of them are laborious and time-consuming. Here we develop a simple and highly sensitive DNA methylation assay based on single base extension reaction and surface enhanced Raman spectroscopy (SERS). In the presence of methylated DNA, gold nanoparticle-modified capture probe can couple with a cyanine 5-deoxyribonucleoside triphosphate (cy5-dGTP) through single base extension reaction, and generates a high SERS signal after further addition of gold nanoparticles to increase the local electromagnetic field. While in the presence of unmethylated DNA, gold nanoparticle-modified capture probe cannot couple with cy5-dGTP due to the presence of a mismatch base, and no SERS signal is observed. This single base extension reaction-based SERS can determine methylated DNA with a detection limit of 3 pM, and can even distinguish as low as 1% methylation level in tumor suppressor gene CDKN2/p16/MTS1 (p16) from the mixtures. Notably, the sensitivity of this assay has improved by 5 orders of magnitude as compared to reported gold nanoparticle-based colorimetric assay, and by 2 orders of magnitude as compared to microarray-based methylation-sensitive single nucleotide primer extension assay (Ms-SNuPE). This method might be further applied to detect the methylation status in tumor-linked genes for cancer diagnosis. (C) 2011 Elsevier B.V. All rights reserved.
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