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Dengue Serotyping with Label-Free DNA Sensor

Abstract

Dengue virus (DENV) is one of the most important mosquito-borne viral diseases in tropical and subtropical regions. Development of severe forms of dengue viral infection such as dengue fever (DF) and dengue hemorrhagic fever (DHF) has claimed many lives. The standard methods of detecting dengue virus are time consuming, laborious, and require skilful personnel. In this study, we propose a method whereby DENV RNA extracted from dengue infected mosquitoes converted into DNA for probe hybridization to generate silver nanocluster strands that could be visualised under UV light. Label-free silver nanocluster based DNA sensors are able to elicit strong fluorescence upon DNA hybridization. Highly specific DNA sequence detection is possible by taking advantage of the specificity of DNA hybridization kinetics. The proposed system is capable of detecting all four dengue DNA serotypes (DENV1-4) without any cross reactivity. A single tube assay format showed better hybridisation efficiency with higher fluorescent intensity generated and a lower detection limit compared to a cocktail probe assay format. The method was able to detect as little as 100nM of amplified double stranded dengue DNA targets using both single and cocktail probes assays. This provides an interesting alternative approach for multiplex DNA sensing utilizing DNA silver nanoclusters as a reporter system.

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Publication details

The article was received on 05 Sep 2017, accepted on 25 Nov 2017 and first published on 30 Nov 2017


Article type: Paper
DOI: 10.1039/C7AY02131C
Citation: Anal. Methods, 2017, Accepted Manuscript
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    Dengue Serotyping with Label-Free DNA Sensor

    S. K. Chan, Y. S. Choong, D. Perera and T. S. Lim, Anal. Methods, 2017, Accepted Manuscript , DOI: 10.1039/C7AY02131C

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