Immunoassay for serodiagnosis of Zika virus infection based on time-resolved Förster resonance energy transfer

PLoS One. 2019 Jul 23;14(7):e0219474. doi: 10.1371/journal.pone.0219474. eCollection 2019.

Abstract

Zika virus (ZIKV) is a mosquito-borne pathogen causing a febrile illness with arthralgia, conjunctivitis and rash. The complications include Guillain-Barré syndrome, congenital brain and other abnormalities and miscarriage. The serodiagnosis of ZIKV infection is hampered by cross-reactivity with other members of the Flavivirus family, notably dengue (DENV). This report describes a novel serological platform for the diagnosis of ZIKV infection. The approach utilizes time-resolved Förster resonance energy transfer (TR-FRET) elicited by two chromophore-labeled proteins (a ZIKV antigen and a super-antigen) simultaneously binding to a given antibody molecule. The antigen used in the assay is ZIKV non-structural protein 1 (NS1) and the super-antigen is bacterial protein L. Three assay variants were developed: the first measuring all anti-ZIKV-NS1 antibodies (LFRET), the second measuring IgM and IgA (acute-LFRET) and the third measuring IgG (immunity-LFRET). The assays were evaluated with a panel of samples from clinical ZIKV cases in travelers (n = 25) and seronegative (n = 24) samples. DENV (n = 38), yellow fever (n = 16) and tick-borne-encephalitis (n = 20) seropositive samples were examined for assessment of flavivirus cross-reactivity. The diagnostic sensitivities of the respective LFRET assays were 92%, 100% and 83%, and the diagnostic specificities 88%, 95% and 100% for LFRET, acute-LFRET and immunity-LFRET. Furthermore, we evaluated the assays against a widely-used commercial ELISA. In conclusion, the new FRET-based serological approaches based on NS1 protein are applicable to diagnosing zika virus infections in travelers and differentiating them from other flavivirus infections.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cohort Studies
  • Enzyme-Linked Immunosorbent Assay
  • Fluorescence Resonance Energy Transfer*
  • Humans
  • Immunoassay / methods*
  • Immunoglobulins / metabolism
  • Serologic Tests*
  • Time Factors
  • Zika Virus / isolation & purification*
  • Zika Virus Infection / blood*
  • Zika Virus Infection / diagnosis*

Substances

  • Immunoglobulins

Grants and funding

The work was mainly funded by a grant from Business Finland, a public funding agency for research in Finland directed by the Finnish Ministry of Employment and the Economy (https://www.businessfinland.fi/). It was further supported by grants from Jane and Aatos Erkko Foundation (https://jaes.fi/), the Academy of Finland (www.aka.fi), the Finnish Medical Society of Finland (https://laaketieteensaatio.fi/) and the Sigrid Jusélius Foundation (https://sigridjuselius.fi/). In addition, several of the researches had individual grants. JH was supported by a grant from the Academy of Finland, SH by grants from the Finnish Foundation for Clinical Chemistry Research (https://www.skky.fi/) and Laboratoriolääketieteen edistämissäätiö (https://www.labes.fi/) and EK by a grant from the Emil Aaltonen Foundation (https://emilaaltonen.fi/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.