High specificity and sensitivity of Zika EDIII-based ELISA diagnosis highlighted by a large human reference panel

PLoS Negl Trop Dis. 2019 Sep 20;13(9):e0007747. doi: 10.1371/journal.pntd.0007747. eCollection 2019 Sep.

Abstract

Background: Zika virus (ZIKV) and Dengue virus (DENV) are often co-endemic. The high protein-sequence homology of flaviviruses renders IgG induced by and directed against them highly cross-reactive against their antigen(s), as observed on a large set of sera, leading to poorly reliable sero-diagnosis.

Methods: We selected Domain III of the ZIKV Envelope (ZEDIII) sequence, which is virus specific. This recombinant domain was expressed and purified for the specific detection of ZEDIII-induced IgG by ELISA from ZIKV-RT-PCR-positive, ZIKV-IgM-positive, flavivirus-positive but ZIKV-negative, or flavivirus-negative sera. We also assessed the reactivity of ZEDIII-specific human antibodies against EDIII of DENV serotype 4 (D4EDIII) as a specific control. Sera from ZEDIII-immunized mice were also tested.

Results: Cross-reactivity of IgG from 5,600 sera against total inactivated DENV or ZIKV was high (71.0% [69.1; 72.2]), whereas the specificity and sensitivity calculated using a representative cohort (242 sera) reached 90% [84.0; 95.8] and 92% [84.5; 99.5], respectively, using a ZEDIII-based ELISA. Moreover, purified human IgG against D2EDIII or D4EDIII did not bind to ZEDIII and we observed no D4EDIII reactivity with ZIKV-induced mouse polyclonal IgGs.

Conclusions: We developed a ZEDIII-based ELISA that can discriminate between past or current DENV and ZIKV infections, allowing the detection of a serological scar from other flaviviruses. This could be used to confirm exposure of pregnant women or to follow the spread of an endemic disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Animals
  • Antibodies, Viral / blood
  • Child
  • Child, Preschool
  • Dengue / diagnosis
  • Dengue / immunology
  • Dengue Virus / immunology
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Humans
  • Immunoglobulin G
  • Infant
  • Male
  • Mice
  • Middle Aged
  • Sensitivity and Specificity
  • Zika Virus / immunology
  • Zika Virus Infection / diagnosis*
  • Zika Virus Infection / immunology
  • Zika Virus Infection / virology

Substances

  • Antibodies, Viral
  • Immunoglobulin G

Grants and funding

This work was partially supported by the European Union’s Horizon 2020 Research and Innovation Programme, under ZIKAlliance Grant Agreement no. 734548, as well as by the European Virus Archive goes global (EVAg) project, under Grant Agreement no. 653316, and partially funded by the Direction Générale de l’Armement and the Service de Santé des Armées, Biomedef PDH-2-NRBC-2-B2-205 and PDH-2-NRBC-2-B-2111. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.