Issue 3, 2018

Colorimetric molecular diagnosis of the HIV gag gene using DNAzyme and a complementary DNA-extended primer

Abstract

We have developed a novel strategy for the colorimetric detection of PCR products by utilizing a target-specific primer modified at the 5′-end with an anti-DNAzyme sequence. A single-stranded DNAzyme sequence folds into a G-quadruplex structure with hemin and shows strong peroxidase activity. When the complementary strand binds to the DNAzyme sequence, it blocks the formation of the G-quadraduplex structure and loses its peroxidase activity. In the presence of the target gene, PCR amplification proceeds, and anti-DNAzyme sequence modified primers present in the reaction mixture form a double strand through primer extension. Therefore, it does not block the DNAzyme sequence. Further, a colorimetric signal is generated by the addition of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) and H2O2 at the end of the reaction. We have successfully detected a single copy of the HIV type 1 gag gene in buffer and 10 copies in human serum. The strategy developed could be used to detect DNA and RNA in complex biological samples by simple primer designing that includes DNAzyme and a DNA extended primer.

Graphical abstract: Colorimetric molecular diagnosis of the HIV gag gene using DNAzyme and a complementary DNA-extended primer

Supplementary files

Article information

Article type
Paper
Submitted
13 Sep 2017
Accepted
16 Dec 2017
First published
19 Dec 2017

Analyst, 2018,143, 695-699

Colorimetric molecular diagnosis of the HIV gag gene using DNAzyme and a complementary DNA-extended primer

S. U. Kim, B. S. Batule, H. Mun, J. Byun, W. Shim and M. Kim, Analyst, 2018, 143, 695 DOI: 10.1039/C7AN01520H

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