Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Observational Study
. 2025 Feb 20;231(2):e337-e344.
doi: 10.1093/infdis/jiae422.

Immunity to Non-Dengue Flaviviruses Impacts Dengue Virus Immunoglobulin G Enzyme-Linked Immunosorbent Assay Specificity in Cambodia

Camila D Odio  1 , Christina Yek  2   3 , Chloe M Hasund  1 , Somnang Man  2 , Piseth Ly  2 , Sreynik Nhek  2 , Sophana Chea  2 , Chanthap Lon  2 , Charlie Voirin  1 , Rekol Huy  4 , Rithea Leang  4 , Chea Huch  4 , Elaine W Lamirande  5 , Stephen S Whitehead  5 , Fabiano Oliveira  2   3 , Jessica E Manning  2   3 , Leah C Katzelnick  1
Affiliations
Observational Study

Immunity to Non-Dengue Flaviviruses Impacts Dengue Virus Immunoglobulin G Enzyme-Linked Immunosorbent Assay Specificity in Cambodia

Camila D Odio et al. J Infect Dis. .

Erratum in

Abstract

Background: Seroprevalence studies are the standard for disease surveillance, and serology determined eligibility for the first dengue vaccine. Expanding flavivirus co-circulation and vaccination complicate testing. We evaluate the accuracy of a common dengue virus serological assay, examine immunity to non-dengue flaviviruses as a contributor to decreased performance, and assess whether alternative cut points may improve assay performance.

Methods: Children (n = 770) aged 2-9 years in Kampong Speu, Cambodia were enrolled in a prospective longitudinal study, and PanBio indirect dengue virus immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) was performed. Plaque reduction neutralization tests (PRNTs) using dengue viruses were performed on a subset to assess the accuracy of the IgG ELISA, and PRNTs with Zika, Japanese encephalitis, and West Nile viruses evaluated immunity to non-dengue flaviviruses. Receiver operating curve analysis identified an alternative cut point to improve IgG ELISA accuracy.

Results: The dengue IgG ELISA had a lower specificity than previously reported (58% vs 93%-100%). Of those with false-positive IgG results, 46% had detectable neutralizing antibodies against other flaviviruses including 14% against West Nile virus. A higher IgG cut point improved the test accuracy in this population.

Conclusions: Physicians and public health authorities should be alert for West Nile in Cambodia. Immunity to non-dengue flaviviruses can impact dengue surveillance.

Clinical trials registration: NCT03534245.

Keywords: West Nile virus; cross-reactivity; dengue virus; seroprevalence; specificity.

PubMed Disclaimer

Conflict of interest statement

Potential conflicts of interest. J. E. M. is now a Clinical Franchise Leader of Influenza in Vaccines Research and Development at Sanofi Pasteur. The work reported in this manuscript was conducted prior to her switching positions. All other authors report no potential conflicts. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed.

Figures

Figure 1.
Figure 1.
Neutralizing antibody titers against JEV, WNV, ZIKV, and DENV in individuals who were (A and C) DENV1–4 naive (ELISA < 0.2) or (B and D) false positive (ELISA >1.1, DENV PRNT50 against all serotypes < 10), or had low DENV nAb (ELISA >1.1, DENV PRNT50 10–20 against ≥1 serotype) as measured by PRNT50 (A and B) and PRNT90 titers (C and D). WNV titers were measured using the chimeric strain for all individuals, and those with nAb against the WNV chimera were confirmed using the wild-type strain. Only positive WNV titers against the wild-type strain are reported. PRNT90 titers were only measured in those with nAb against ≥2 nondengue flaviviruses. Abbreviations: DENV, dengue virus; ELISA, enzyme-linked immunosorbent assay; JEV, Japanese encephalitis virus; nAb, neutralizing antibody; PRNT, plaque reduction neutralization test; WNV, West Nile virus; ZIKV, Zika virus.
Figure 2.
Figure 2.
Receiver operating curve analysis for all individuals with ELISA and PRNT testing (n = 336). Sensitivity and specificity are reported for ELISA cut points of 1.1 versus 1.59. Abbreviations: CI, confidence interval; ELISA, enzyme-linked immunosorbent assay; PRNT, plaque reduction neutralization test.
See this image and copyright information in PMC

Update of

References

    1. Postler TS, Beer M, Blitvich BJ, et al. Renaming of the genus Flavivirus to Orthoflavivirus and extension of binomial species names within the family Flaviviridae. Arch Virol 2023; 168:224. - PubMed
    1. Du M, Jing W, Liu M, Liu J. The global trends and regional differences in incidence of dengue infection from 1990 to 2019: an analysis from the global burden of disease study 2019. Infect Dis Ther 2021; 10:1625–43. - PMC - PubMed
    1. Pierson TC, Diamond MS. The continued threat of emerging flaviviruses. Nat Microbiol 2020; 5:796–812. - PMC - PubMed
    1. Chan KR, Ismail AA, Thergarajan G, et al. Serological cross-reactivity among common flaviviruses. Front Cell Infect Microbiol 2022; 12:975398. - PMC - PubMed
    1. Strategic Advisory Group of Experts on Immunization . Background paper on dengue vaccines. https://terrance.who.int/mediacentre/data/sage/SAGE_Docs_Ppt_Apr2016/10_.... Accessed 17 November 2023.

Publication types

MeSH terms

Associated data