Competitive ELISA for a serologic test to detect dengue serotype-specific anti-NS1 IgGs using high-affinity UB-DNA aptamers
Serologic tests to detect specific IgGs to antigens related to viral infections are urgently needed for diagnostics and therapeutics. We present a diagnostic method for serotype-specific IgG identification of dengue infection by a competitive enzyme-linked immunosorbent assay (ELISA), using high-aff...
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sg-ntu-dr.10356-1537982023-03-05T16:50:38Z Competitive ELISA for a serologic test to detect dengue serotype-specific anti-NS1 IgGs using high-affinity UB-DNA aptamers Matsunaga, Ken-Ichiro Kimoto, Michiko Lim, Vanessa Weixun Thein, Tun-Linn Vasoo, Shawn Leo, Yee Sin Sun, William Hirao, Ichiro Lee Kong Chian School of Medicine (LKCMedicine) National Centre for Infectious Diseases Department of Infectious Diseases, Tan Tock Seng Hospital Science::Medicine Biological Techniques Diseases Serologic tests to detect specific IgGs to antigens related to viral infections are urgently needed for diagnostics and therapeutics. We present a diagnostic method for serotype-specific IgG identification of dengue infection by a competitive enzyme-linked immunosorbent assay (ELISA), using high-affinity unnatural-base-containing DNA (UB-DNA) aptamers that recognize the four categorized serotypes. Using UB-DNA aptamers specific to each serotype of dengue NS1 proteins (DEN-NS1), we developed our aptamer-antibody sandwich ELISA for dengue diagnostics. Furthermore, IgGs highly specific to DEN-NS1 inhibited the serotype-specific NS1 detection, inspiring us to develop the competitive ELISA format for dengue serotype-specific IgG detection. Blood samples from Singaporean patients with primary or secondary dengue infections confirmed the highly specific IgG detection of this format, and the IgG production initially reflected the serotype of the past infection, rather than the recent infection. Using this dengue competitive ELISA format, cross-reactivity tests of 21 plasma samples from Singaporean Zika virus-infected patients revealed two distinct patterns: 8 lacked cross-reactivity, and 13 were positive with unique dengue serotype specificities, indicating previous dengue infection. This antigen-detection ELISA and antibody-detection competitive ELISA combination using the UB-DNA aptamers identifies both past and current viral infections and will facilitate specific medical care and vaccine development for infectious diseases. Agency for Science, Technology and Research (A*STAR) National Research Foundation (NRF) Published version This work was supported by the Institute of Bioengineering and Bioengineering (Biomedical Research Council, Agency for Science, Technology and Research, Singapore) and the National Research Foundation of Singapore (NRF-CRP17-2017-07). 2021-12-29T02:39:29Z 2021-12-29T02:39:29Z 2021 Journal Article Matsunaga, K., Kimoto, M., Lim, V. W., Thein, T., Vasoo, S., Leo, Y. S., Sun, W. & Hirao, I. (2021). Competitive ELISA for a serologic test to detect dengue serotype-specific anti-NS1 IgGs using high-affinity UB-DNA aptamers. Scientific Reports, 11(1), 18000-. https://dx.doi.org/10.1038/s41598-021-97339-8 2045-2322 https://hdl.handle.net/10356/153798 10.1038/s41598-021-97339-8 34504185 2-s2.0-85114747018 1 11 18000 en NRF-CRP17-2017-07 Scientific Reports © 2021 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. application/pdf |
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Science::Medicine Biological Techniques Diseases Matsunaga, Ken-Ichiro Kimoto, Michiko Lim, Vanessa Weixun Thein, Tun-Linn Vasoo, Shawn Leo, Yee Sin Sun, William Hirao, Ichiro Competitive ELISA for a serologic test to detect dengue serotype-specific anti-NS1 IgGs using high-affinity UB-DNA aptamers |
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Serologic tests to detect specific IgGs to antigens related to viral infections are urgently needed for diagnostics and therapeutics. We present a diagnostic method for serotype-specific IgG identification of dengue infection by a competitive enzyme-linked immunosorbent assay (ELISA), using high-affinity unnatural-base-containing DNA (UB-DNA) aptamers that recognize the four categorized serotypes. Using UB-DNA aptamers specific to each serotype of dengue NS1 proteins (DEN-NS1), we developed our aptamer-antibody sandwich ELISA for dengue diagnostics. Furthermore, IgGs highly specific to DEN-NS1 inhibited the serotype-specific NS1 detection, inspiring us to develop the competitive ELISA format for dengue serotype-specific IgG detection. Blood samples from Singaporean patients with primary or secondary dengue infections confirmed the highly specific IgG detection of this format, and the IgG production initially reflected the serotype of the past infection, rather than the recent infection. Using this dengue competitive ELISA format, cross-reactivity tests of 21 plasma samples from Singaporean Zika virus-infected patients revealed two distinct patterns: 8 lacked cross-reactivity, and 13 were positive with unique dengue serotype specificities, indicating previous dengue infection. This antigen-detection ELISA and antibody-detection competitive ELISA combination using the UB-DNA aptamers identifies both past and current viral infections and will facilitate specific medical care and vaccine development for infectious diseases. |
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Lee Kong Chian School of Medicine (LKCMedicine) |
author_facet |
Lee Kong Chian School of Medicine (LKCMedicine) Matsunaga, Ken-Ichiro Kimoto, Michiko Lim, Vanessa Weixun Thein, Tun-Linn Vasoo, Shawn Leo, Yee Sin Sun, William Hirao, Ichiro |
format |
Article |
author |
Matsunaga, Ken-Ichiro Kimoto, Michiko Lim, Vanessa Weixun Thein, Tun-Linn Vasoo, Shawn Leo, Yee Sin Sun, William Hirao, Ichiro |
author_sort |
Matsunaga, Ken-Ichiro |
title |
Competitive ELISA for a serologic test to detect dengue serotype-specific anti-NS1 IgGs using high-affinity UB-DNA aptamers |
title_short |
Competitive ELISA for a serologic test to detect dengue serotype-specific anti-NS1 IgGs using high-affinity UB-DNA aptamers |
title_full |
Competitive ELISA for a serologic test to detect dengue serotype-specific anti-NS1 IgGs using high-affinity UB-DNA aptamers |
title_fullStr |
Competitive ELISA for a serologic test to detect dengue serotype-specific anti-NS1 IgGs using high-affinity UB-DNA aptamers |
title_full_unstemmed |
Competitive ELISA for a serologic test to detect dengue serotype-specific anti-NS1 IgGs using high-affinity UB-DNA aptamers |
title_sort |
competitive elisa for a serologic test to detect dengue serotype-specific anti-ns1 iggs using high-affinity ub-dna aptamers |
publishDate |
2021 |
url |
https://hdl.handle.net/10356/153798 |
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1759855767266000896 |