95% Interval Confidence (IC) value is also indicated and showed in dotted lines above and under the linear correlation. was performed, and sera samples were collected for IgM/IgG detection are indicated in figures.(TIF) pntd.0008039.s002.tif (259K) GUID:?865F3011-E707-4505-8449-81A654C8A904 S3 Fig: rNS1-based ELISA test for the detection of IgM/IgG antibodies against DENV1-4 rNS1 6xHis-tagged proteins. A to H) Detection of IgM/IgG antibodies from 4 different immunized mice with NS1 V5-tagged create for each DENV serotype (DENV-1, DENV-2, DENV-3, and DENV-4). Plates were coated with purified DENV1-rNS1, Phen-DC3 DENV2-rNS1, DENV3-rNS1 and DENV4-rNS1 antigens for the detection of antibodies from mice immunized with each DENV serotype, respectively. * each ELISA result includes the average of two biological replicates.(TIF) pntd.0008039.s003.tif (1.0M) GUID:?264DA090-F283-4C05-9D50-7B6A1EFC264D S1 Table: Optimal cut-off ideals of P/N percentage for IgM/IgG antibody detection from sera of immunized mice and infected individuals. P/N: positive to the bad percentage (OD450 of test specimen divided from the mean OD450 of bad control specimens). ROC: comparative receiver operating characteristic curve rNS1: recombinant non-structural protein 1 TBEV: Tick-borne encephalitis computer virus WNV: Western Nile computer virus ZIKV: Zika computer virus DENV1-4: Dengue computer virus serotype 1, serotype 2, serotype 3 and serotype 4.(TIF) pntd.0008039.s004.tif (69K) GUID:?C1FAB182-E0CF-4B37-949B-4ECA11C8C3E5 S2 Table: Quantity of positive samples from TBEV infected patients. TBEV: Tick-borne encephalitis computer virus ELISA: enzyme-linked immunosorbent assay rNS1: recombinant non-structural protein 1 a A total of 11 samples were tested for each group, 1st (1st) and second (2nd) phase of TBEV illness b A total of 34 samples were tested for the acute phase of TBEV illness c A total of 44 samples were tested for the convalescent phase of TBEV illness d plates were coated with inactivated TBEV E antigen, Enzygnost; Simens GmbH e plates were coated with recombinant non-structural protein 1 of TBEV.(TIF) pntd.0008039.s005.tif (78K) GUID:?788C4913-F2C2-4B59-AC28-1DF1849A80CC S3 Table: Quantity of positive samples from flavivirus infected patients. WNV: Western Nile computer virus ZIKV: Zika computer virus DENV1-4: Dengue computer virus serotype 1, serotype 2, serotype 3 and serotype 4 ELISA: enzyme-linked immunosorbent assay rNS1: recombinant non-structural protein 1 a IgM/IgG antibodies were detected by using commercial kits according to the manufacturers instructions for WNV (plates coated with recombinant WNV E antigen, Focus Diagnostics), ZIKV (plates coated with ZIKV recombinant NS1 antigen, Euroimmune; Labordiagnostika AG) and, DENV1-4 (plates coated with DENV type 2 E antigen, NovaTec, immunodiagnostic GmbH). b IgM/IgG antibodies were recognized using purified rNS1 proteins TNFRSF1A of WNV, ZIKV and all DENV serotypes.(TIF) pntd.0008039.s006.tif (77K) GUID:?75B85124-68C6-4CFF-A0EE-1766C160D569 S4 Table: Quantity of TBEV positive IgM/IgG samples out of the total number of samples analyzed by commercial ELISA and rNS1-based ELISA assays. TBEV: Tick-borne encephalitis computer virus ELISA: enzyme-linked immunosorbent assay rNS1: recombinant non-structural protein 1 VBT: vaccine breakthrough NA: non-available a IgM/IgG antibodies were detected by using a commercial kit according to the manufacturers instructions for TBEV (plates were coated with inactivated TBEV E antigen, Enzygnost; Simens GmbH b IgM/IgG antibodies were recognized by rNS1-centered ELISA covering the plates with the rNS1 protein of TBEV.(TIF) pntd.0008039.s007.tif (78K) GUID:?463C8AFF-3EA9-4952-9BB0-0EDA4761BA41 Attachment: Submitted filename: value of 0.0001 rejected the null hypothesis that there is present no correlation between commercial and rNS1-based ELISA methods. 95% Interval Confidence (IC) value is also indicated and showed in dotted lines above and under the linear correlation. 100 sera samples (n) from different phases of TBEV illness were included in the analysis. * each ELISA result includes the average of two biological replicates. Based on 89 IgM/IgG positive samples (100% level of sensitivity) and 11 IgM/IgG bad samples (100% specificity) analyzed by commercial Phen-DC3 ELISA, we identified the level of sensitivity and specificity of the TBEV rNS1-centered ELISA with 95% confidence intervals (CI). The level of sensitivity was 91% and 100%, while the specificity was 100% and 91% for IgM (95% CI, 0.77 to 0.89) and IgG (95% CI, 0.62 to 0.81) detection, respectively (Fig 3E and 3F). Combined IgM/IgG level of sensitivity and specificity was 96% Phen-DC3 among the 100 TBEV-positive specimens tested. rNS1-centered ELISA for WNV, ZIKV and DENV 1C4 infections A cohort of 84 RT-PCR-positive specimens from infected individuals with flavivirus (travelers firstly.