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

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — UB-DNA aptamers recognize each DEN1-NS1 variant. (A) Differences in the amino acid sequences of DEN1-NS1 proteins derived from the clinical samples (PD1-1, PD1-2 and PD1-3) and the original DEN1-NS1 target from The Native Antigen Company (NAD1). The different amino acids between DEN1-NS1 variant 2 (from PD1-2/PD1-3) and DEN1-NS1 variant 2 from PD1-1 are indicated in red. The sequence identities are presented as compared to NAD1 (left column) and PD1-2/1-3 (right column). (B) Presumed secondary structures of UB-DNA aptamers, AptD1 and AptD1b, which specifically target the original DEN1-NS1 variant 1 (NAD1) and a recombinant DEN1-NS1 variant 2 protein (from PD1-2/PD1-3), respectively. The kinetic binding parameters, dissociation constant (K_D), of each aptamer to each cognate DEN1-NS1 variant were determined by an SPR analysis (Supplementary Figure S14). (C) DEN-NS1 detection by the aptamer−antibody (Ab#D06) ELISA formats using AptD1 (D1-1-48h) and AptD1b (19D1F1-3). As the test samples (10 μl), different clinical sera (PD1-1, PD1-2 and PD1-3) and each serotype DEN-NS1 recombinant protein from The Native Antigen Company (NAD1, NAD2, NAD3, and NAD4; 100 ng/ml) were used.