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Open AccessArticle

The Dengue ED3 Dot Assay, a Novel Serological Test for the Detection of Denguevirus Type-Specific Antibodies and Its Application in a Retrospective Seroprevalence Study

1
Department of Virology, Bernhard Nocht Institute for Tropical Medicine, Bernhard Nocht Str. 74, 20359 Hamburg, Germany
2
Virology Unit, Institut Pasteur in Cambodia, 5 Monivong Boulevard, 12201 Phnom Penh, Cambodia
3
GlaxoSmithKline, Vaccines R&D, 23 Rochester Park, Singapore 139234, Singapore
*
Author to whom correspondence should be addressed.
These authors contributed equally to this article.
Viruses 2019, 11(4), 304; https://doi.org/10.3390/v11040304
Received: 6 February 2019 / Revised: 22 March 2019 / Accepted: 25 March 2019 / Published: 27 March 2019
(This article belongs to the Section Animal Viruses)
There are four distinct antigenic serotypes of dengue viruses (DENV-1-4). Sequential infections with different serotypes lead to cross-reactive but also serotype-specific neutralizing antibody responses. Neutralization assays are considered as gold standard for serotype-specific antibody detection. However, for retrospective seroprevalence studies, access to large serum quantities is limited making neutralization assays well-nigh impossible. Therefore, a serological test, wasting only 10 µL serum, was developed using fusion proteins of maltose binding protein and E protein domain 3 (MBP-ED3) as antigens. Twelve MBP-ED3 antigens for DENV-1-4, three MBP-ED3 antigens for WNV, JEV, and TBEV, and MBP were dotted onto a single nitrocellulose strip. ED3 dot assay results were compared to virus neutralization and ED3 ELISA test results, showing a >90% accordance for DENV-1 and a 100% accordance for DENV-2, making the test specifically useful for DENV-1/-2 serotype-specific antibody detection. Since 2010, DENV-1 has replaced DENV-2 as the dominant serotype in Cambodia. In a retrospective cohort analysis, sera collected during the DENV-1/-2 endemic period showed a shift to DENV-2-specific antibody responses in 2012 paralleled by the decline of DENV-2 infections. Altogether, the ED3 dot assay is a serum-, time- and money-saving diagnostic tool for serotype-specific antibody detection, especially when serum samples are limited. View Full-Text
Keywords: dengue virus; serology; serotype; antigen assay; domain 3; E protein; seroprevalence dengue virus; serology; serotype; antigen assay; domain 3; E protein; seroprevalence
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Auerswald, H.; Klepsch, L.; Schreiber, S.; Hülsemann, J.; Franzke, K.; Kann, S.; Y, B.; Duong, V.; Buchy, P.; Schreiber, M. The Dengue ED3 Dot Assay, a Novel Serological Test for the Detection of Denguevirus Type-Specific Antibodies and Its Application in a Retrospective Seroprevalence Study. Viruses 2019, 11, 304.

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