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

Heidi Auerswald¹  Janne Hülsemann¹  Sebastian Schreiber¹  Kati Franzke¹  Michael Schreiber¹  Leonard Klepsch¹  Simone Kann¹  Bunthin Y²  Philippe Buchy²  Veasna Duong² 

[1] Department of Virology, Bernhard Nocht Institute for Tropical Medicine, Bernhard Nocht Str. 74, 20359 Hamburg, Germany;Virology Unit, Institut Pasteur in Cambodia, 5 Monivong Boulevard, 12201 Phnom Penh, Cambodia;

关键词: dengue virus;    serology;    serotype;    antigen assay;    domain 3;    E protein;    seroprevalence;   

DOI  :  10.3390/v11040304

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown