Design, development, and validation of multi-epitope proteins for serological diagnosis of Zika virus infections and discrimination from dengue virus seropositivity

PLoS Negl Trop Dis. 2024 Apr 18;18(4):e0012100. doi: 10.1371/journal.pntd.0012100. eCollection 2024 Apr.

Abstract

Zika virus (ZIKV), an arbovirus from the Flaviviridae family, is the causative agent of Zika fever, a mild and frequent oligosymptomatic disease in humans. Nonetheless, on rare occasions, ZIKV infection can be associated with Guillain-Barré Syndrome (GBS), and severe congenital complications, such as microcephaly. The oligosymptomatic disease, however, presents symptoms that are quite similar to those observed in infections caused by other frequent co-circulating arboviruses, including dengue virus (DENV). Moreover, the antigenic similarity between ZIKV and DENV, and even with other members of the Flaviviridae family, complicates serological testing due to the high cross-reactivity of antibodies. Here, we designed, produced in a prokaryotic expression system, and purified three multiepitope proteins (ZIKV-1, ZIKV-2, and ZIKV-3) for differential diagnosis of Zika. The proteins were evaluated as antigens in ELISA tests for the detection of anti-ZIKV IgG using ZIKV- and DENV-positive human sera. The recombinant proteins were able to bind and detect anti-ZIKV antibodies without cross-reactivity with DENV-positive sera and showed no reactivity with Chikungunya virus (CHIKV)- positive sera. ZIKV-1, ZIKV-2, and ZIKV-3 proteins presented 81.6%, 95%, and 66% sensitivity and 97%, 96%, and 84% specificity, respectively. Our results demonstrate the potential of the designed and expressed antigens in the development of specific diagnostic tests for the detection of IgG antibodies against ZIKV, especially in regions with the circulation of multiple arboviruses.

MeSH terms

  • Antibodies, Viral
  • Arboviruses*
  • Chikungunya Fever*
  • Dengue Virus*
  • Dengue*
  • Epitopes
  • Humans
  • Immunoglobulin G
  • Zika Virus Infection* / diagnosis
  • Zika Virus* / genetics

Substances

  • Epitopes
  • Antibodies, Viral
  • Immunoglobulin G

Grants and funding

FGF received funds from Fundação de Amparo à Pesquisa do Estado de Minas Gerais - FAPEMIG (grant numbers APQ-03081-17, APQ-04295-17 and RED-00282-16); and from Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq (grant # 402256/2022-0). The study was also financed by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES) – Finance Code 001, through fellowships given to post-graduation students involved in the work. FGF, MLN, JGACR and JSA are CNPQ-PQ fellowship recipients. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.