Skip to main content
. 2021 Jun 25;49(20):11407–11424. doi: 10.1093/nar/gkab515

Figure 3.

Figure 3.

UB-DNA aptamers, D1-1-48h (AptD1), D2-1d-72h (AptD2), D3-2-59h (AptD3), and D4-3-57h (AptD4), specifically recognize the targeted DEN-NS1, allowing for serotype-specific dengue diagnostics. (A) Electrophoresis gel-mobility shift assay (EMSA) of the aptamer−NS1 complex formation using each UB-DNA aptamer and their variants without unnatural bases. (B) Binding analysis of the UB-DNA aptamers to each DEN-NS1 by a Biacore T200 SPR system at 25°C. Each aptamer's kinetic binding parameters, dissociation constant (KD), and association and dissociation rates (kon and koff) to each targeted DEN-NS1 were determined. (C) Serotype-specific DEN-NS1 detection by ELISA in combination with the UB-DNA aptamer and antibody (Ab#D06) pair. In ELISA, a 10-μl portion of a 100 ng/ml solution of each flavivirus NS1 protein (DENV serotypes 1–4 NS1 proteins, and Zika virus NS1 proteins of a Brazilian strain and a Ugandan strain, purchased from The Native Antigen Company) was used in buffer. The sample size is two per each combination set, and the error bars represent one average deviation. The bars with wavy lines indicate that the signal in at least one of the two sample wells was saturated (OD450 > 4.000).