Wells. 4.10. Anti-DENV Complement-Fixing Antibody Assay Traits The limit of detection (LOD), limit of quantitation (LLOQ), linearity, and precision are described in the supplemental material. LOD and LLOQ had been determined per Quinn et al. [64]. four.11. Lurasidone-d8 Technical Information Homology of Flavivirus Envelope Protein Various sequence alignment and phylogenetic tree analysis with the envelope amino acid sequence of DENV1 (strain Western Pacific; GenBank accession No. AAN06981.1), DENV2 (strain New Guinea C; GenBank accession No. AAP73862.1), DENV3 (strain Sleman/78; GenBank accession No. AAT69740.1), or DENV4 (strain 1228; GenBank accession No. AER00190.1), Zika virus (strain PRVABC59; GenBank accession No. AMZ03556.1), yellow fever virus (strain Stamaril, Sanofi Pasteur, Lyon, France; GenBank accession No. QGN18661.1), West Nile virus (strain NY99; GenBank accession No. AAG02038.1), Japanese encephalitis virus (strain SA 14-14-2; GenBank accession No. ARE67893.1), and tick-borne encephalitis virus (strain K23; GenBank accession No. AAC62093.1) have been carried out applying web interface for Clustal Omega and protein blast accessible at https: //www.ebi.ac.uk/Tools/msa/clustalo/ (accessed on 19 Could 2021) and https://blast.ncbi. nlm.nih.gov/Blast.cgi# (accessed on 19 Might 2021), respectively. The former was applied to construct a phylogenetic tree, though the latter, to calculate protein % identity. four.12. Statistical Analysis Specificity of anti-DENV complement-fixing antibodies were calculated depending on the percentage of samples with MNT50 10 (seronegative) that were beneath the threshold titer of 3 EU/mL. On the other hand, sensitivity was calculated as the percentage of samples with MNT50 ten (seropositive) that have been equal to or above the threshold titer of 3 EU/mL. The receiver operating characteristic (ROC) curves had been made by plotting the accurate seropositive price (defined as the price of complement-fixing antibodies with titers equal to or above the threshold titer of 3 EU/mL in seropositive samples (MNT50 ten)) against the false seropositive rate (defined as complement-fixing antibody titers equal or above the threshold titer of 3 EU/mL in samples regarded as seronegative (MNT50 ten)) at numerous complement-fixing antibody thresholds from three EU/mL to roughly 1000 EU/mL. The region beneath the ROC was calculated as a single numerical measurement to describe the efficiency of your complement-fixing antibody titers as a biomarker to classify dengue serostatus. Correlation analysis among complement-fixing antibody titers and microneutralization titers or total binding IgG concentrations was carried out Etomidate-d5 supplier employing log10 -transformed values employing JMP version 13.1.0 application (SAS Institute Inc., San Diego, CA, USA). Samples were color-coded based on DENV serostatus (blue–seronegative; red–seropositive). Correlation evaluation involving complement-fixing and C3d deposition antibody concentrations was carried out making use of log10 -transformed values employing GraphPad Prism version eight.1.0 (GraphPad Software, Inc., San Diego, CA, USA).Int. J. Mol. Sci. 2021, 22,16 ofSupplementary Components: The following are obtainable on the internet at mdpi/article/10 .3390/ijms222112004/s1. Author Contributions: Conceptualization, E.J.M.N. and H.J.D.; methodology, E.J.M.N., R.B., E.K.; formal analysis, E.J.M.N., B.N., A.P., R.B.; E.K. and H.J.D.; investigation, E.J.M.N., B.N., A.P., R.B., E.K. and H.J.D.; data curation, E.J.M.N., B.N., A.P.; writing–original draft preparation, E.J.M.N., B.N., A.P., R.B., E.K. and.