RESUMO
The dengue hemorrhagic fever or dengue shock syndrome has become a severe human fatal disease caused by infection with one of the four closely related but serologically distinct dengue viruses (DENVs). All four dengue serotypes are currently co-circulating throughout the subtropics and tropics. Since the fatality rate increases severely when a secondary infection occurs by a virus serotype different from that of the initial infection, serotype identification is equally important as virus detection. In this study, the development and validation of a rapid and quantitative DENV serotype-specific (serotypes 1-4) biosensor are reported by optimizing the stable system between cadmium selenide tellurium sulphide fluorescent quantum dots (CdSeTeS QDs) and gold nanoparticles (AuNPs). Four different nanoprobes are designed using each primer-probe serotype-specific hairpin single-stranded DNA covalently bound at different positions to CdSeTeS QDs, which generates an altered fluorescence signal for each serotype of DENV. In fourplex reactions with free functionalized AuNPs and the four nanoprobes, the standard dilutions of the target virus DNA from 10-15 to 10-10 M were successfully detected. The limit of detection was found to be in the femtomolar range for all four serotypes, where the serotype detection ability was undoubtedly established. To confirm the applicability of this sensing performance in long chained complex RNAs, the sensor was also applied successfully to RNAs extracted from DENV culture fluids for serotype identification as well as quantification, which can lead to a potential diagnostic probe for point-of-care detection.
