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Fig 1.

The LFRET assay principle.

The Eu-labeled antigen (ZIKV NS1), Alexa-Fluor-labeled protein L and the serum sample are mixed together in a 384-well microplate. The former binds to the light chains of the antibodies, which if ZIKV-NS1-specific also bind the europium-labeled NS1 antigen. The mixture is exposed to UV light and the close proximity allows the donor and acceptor to elicit the FRET signal, which is measured.

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Fig 1 Expand

Table 1.

Reference test results for ZIKV-positive samples.

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Fig 2.

Class-independent LFRET scores for the first cohort of samples.

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Table 2.

Class-independent LFRET test outcomes in different sample categories.

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Table 2 Expand

Fig 3.

Class-independent LFRET scores measured from the full panel of ZIKV-positive, heterologous flavivirus-positive (DENV, YFV or TBEV) and flavivirus-negative samples.

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Fig 3 Expand

Table 3.

Test outcomes in class-specific LFRET tests compared with the reference tests.

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Fig 4.

a) Acute-LFRET scores of IgM-positive and -negative samples b) Immunity-LFRET scores of IgG-positive and -negative samples.

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Table 4.

Test outcomes when using non-rigorous sample inclusion criteria.

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Fig 5.

Euroimmun ELISA vs. class-specific LFRET.

a) Optical density values in Euroimmun IgM ELISA compared to the acute-LFRET scores of the same samples; b) OD values in Euroimmun IgG ELISA test compared to the immunity-LFRET score.

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Fig 5 Expand

Table 5.

Comparison of the outcomes of the Euroimmun ELISA test with class-specific LFRET test outcomes.

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Table 5 Expand

Table 6.

The samples with inconsistent results between Euroimmun ELISA and class-specific LFRET assay.

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