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Comparative Study
. 2024 May 15;16(5):788.
doi: 10.3390/v16050788.

Comparison of Five Serological Methods for the Detection of West Nile Virus Antibodies

Affiliations
Comparative Study

Comparison of Five Serological Methods for the Detection of West Nile Virus Antibodies

Philipp Girl et al. Viruses. .

Abstract

The West Nile Virus (WNV), a member of the family Flaviviridae, is an emerging mosquito-borne flavivirus causing potentially severe infections in humans and animals involving the central nervous system (CNS). Due to its emerging tendency, WNV now occurs in many areas where other flaviviruses are co-occurring. Cross-reactive antibodies with flavivirus infections or vaccination (e.g., tick-borne encephalitis virus (TBEV), Usutu virus (USUV), yellow fever virus (YFV), dengue virus (DENV), Japanese encephalitis virus (JEV)) therefore remain a major challenge in diagnosing flavivirus infections. Virus neutralization tests are considered as reference tests for the detection of specific flavivirus antibodies, but are elaborate, time-consuming and need biosafety level 3 facilities. A simple and straightforward assay for the differentiation and detection of specific WNV IgG antibodies for the routine laboratory is urgently needed. In this study, we compared two commercially available enzyme-linked immunosorbent assays (anti-IgG WNV ELISA and anti-NS1-IgG WNV), a commercially available indirect immunofluorescence assay, and a newly developed in-house ELISA for the detection of WNV-NS1-IgG antibodies. All four tests were compared to an in-house NT to determine both the sensitivity and specificity of the four test systems. None of the assays could match the specificity of the NT, although the two NS1-IgG based ELISAs were very close to the specificity of the NT at 97.3% and 94.6%. The in-house WNV-NS1-IgG ELISA had the best performance regarding sensitivity and specificity. The specificities of the ELISA assays and the indirect immunofluorescence assays could not meet the necessary specificity and/or sensitivity.

Keywords: WNV; anti-NS1-IgG WNV; flavivirus; mosquito-borne; neutralization test.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Proportionate distribution of sera determined as positive or negative in IIFA for negative and positive neutralization test (NT) results.
Figure 2
Figure 2
Distribution of Vienna units (VIEU)/mL determined by ELISA (A) and NS1 ELISA (B) or OD determined by IH-NS1 ELISA (C) for negative and positive neutralization test results (NT). The cut-off of each assay is shown as a red dashed line.

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