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

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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

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Publication details

The article was received on 13 Sep 2017, accepted on 16 Dec 2017 and first published on 19 Dec 2017


Article type: Paper
DOI: 10.1039/C7AN01520H
Citation: Analyst, 2018, Advance Article
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    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, Advance Article , DOI: 10.1039/C7AN01520H

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