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

Jessica Denis (1, 2) , Sarah Attoumani (3, 4) , Patrick Gravier (4, 2) , Bernard Tenebray (4, 2) , Annabelle Garnier (1) , Sébastien Briolant (5, 6) , Franck de Laval (7, 8) , Véronique Chastres (9) , Gilda Grard (4, 2) , Isabelle Leparc-Goffart (4, 2) , Bruno Coutard (4, 3) , Cyril Badaut (1, 2)

1 Unité de Biothérapies anti-Infectieuses et Immunité [Brétigny-sur-Orge]
2 Centre National de Référence des Arbovirus [Marseille]
3 AFMB - Architecture et fonction des macromolécules biologiques
4 UVE - Unité des Virus Emergents
5 Unité de Parasitologie et d’Entomologie
6 VITROME - Vecteurs - Infections tropicales et méditerranéennes
7 Centre d'Epidémiologie et de Santé Public des Armées
8 SESSTIM - U1252 INSERM - Aix Marseille Univ - UMR 259 IRD - Sciences Economiques et Sociales de la Santé & Traitement de l'Information Médicale
9 IRBA - Institut de Recherche Biomédicale des Armées [Brétigny-sur-Orge]

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.

Domaines: Virologie

Jessica Denis, Sarah Attoumani, Patrick Gravier, Bernard Tenebray, Annabelle Garnier, et al.. High specificity and sensitivity of Zika EDIII-based ELISA diagnosis highlighted by a large human reference panel. PLoS Neglected Tropical Diseases, 2019, 13 (9), pp.e0007747. ⟨10.1371/journal.pntd.0007747⟩. ⟨inserm-02416987⟩