Article Figures & Data
Figures
- Figure 1 Flowchart showing phases I and II of this study
Center 1 (Innsbruck) performed 5 assays (live CBA-IF MOG-IgG (H + L), live CBA-IF MOG-IgG(Fc), live CBA-FACS MOG-IgG(Fc), live CBA-IF MOG-IgM, and ELISA MOG-IgG); center 2 (Mayo Clinic) performed 1 assay (live CBA-FACS MOG-IgG1); center 3 (Oxford) performed 2 assays (live CBA-IF MOG-IgG (H + L) and live CBA-IF MOG-IgG1); center 4 (Sydney) performed 1 assay (live CBA-FACS MOG-IgG (H + L)), which was repeated twice; center 5 (Euroimmun) performed 2 assays (fixed CBA-IF MOG-IgG(Fc) and ELISA MOG-IgG(Fc)). CBA = cell-based assay; FACS = fluorescence-activated cell sorting; IF = immunofluorescence; IfQ = Institute for Quality Assurance; Ig = immunoglobulin; MOG = myelin oligodendrocyte glycoprotein.
- Figure 2 Agreement of MOG-Ab assays on clear positive and negative samples
(A) Heatmap of the qualitative results for samples sent as clearly positive (n = 39) or negative (n = 40). Each column is an individual assay (1–8 MOG-IgG CBAs, 9 MOG-IgM CBAs, and 10–11 MOG-IgG ELISAs), and each row is an individual serum sample. Results are based on qualitative results; negative samples are black, and positive samples are red. The samples are shown according to their serostatus sent by the individual centers. (B) Agreement of MOG-IgG CBAs according to the samples sent (pos = positive, neg = negative). Results (in % of all samples) are grouped according to their agreement in all 8 CBAs or in the 7 live CBAs (red: positive in all CBAs, black: negative in all CBAs, white: discordant). (C) Quantitative results for all assays. The cutoff values for all assays except assay Nr. 7 are indicated by the dashed gray lines. For assay Nr. 7, cutoff levels for pediatric samples (blue dots) are indicated by the blue dashed line, and cutoff levels for adult samples (red dots) are indicated by the red dashed line. The quantitative range of each assay result for its probability to be seropositive in all live CBAs is indicated by the dotted line and shaded in darker gray (100% probability), whereas the range of discordant samples is shaded in light gray. A single sample with high IgM titer 1:5120 and low positive in the IgG (H + L) and on IgG1 but not in another IgG1 and the IgG(Fc) assays is indicated by the larger white dot. BR = binding ratio; CBA = cell-based assay; dMFI = delta mean fluorescence intensity; Ig = immunoglobulin; MOG = myelin oligodendrocyte glycoprotein.
- Figure 3 Correlation of MOG-IgG assays and reproducibility of assay results
(A) Heatmap of Spearman correlation coefficients for all correlations. (B) Correlation of illustrative live (Nr. 2, 4, 6, and 7) and fixed CBAs (Nr. 8): Nr. 2 (center 1): CBA-IF IgG(Fc) titer (1), Nr. 4 (center 2): CBA-FACS IgG1 binding ratio, Nr. 6 (center 3): CBA-IF IgG1 binding score, Nr. 7 (center 4): CBA-FACS IgG (H + L) delta mean fluorescence intensity, Nr, 8 (center 5): CBA-IF IgG(Fc) titer (1). The cutoff values for all assays except assay Nr. 7 are indicated by the dashed gray lines. For assay Nr. 7, cutoff levels for pediatric samples (blue dots) are indicated by the blue dashed line, and cutoff levels for adult samples (red dots) are indicated by the red dashed line. Samples that are positive in live CBAs but not the fixed CBA (assay Nr. 8) are indicated by the white dots. (C) Qualitative results for 9 positive and 9 negative samples from phase I retested in a blinded way by all assays in phase II. CBA = cell-based assay; IF = immunofluorescence; Ig = immunoglobulin; MOG = myelin oligodendrocyte glycoprotein.
- Figure 4 Agreement of MOG-Ab assays on low positive and borderline negative samples and blood donors
Qualitative and quantitative results of all MOG-IgG CBAs for blood donors (BD, n = 30), low positive (n = 39), and borderline negative (n = 40) samples. (A) Qualitative results according to the serostatus sent. Each column is an individual MOG-IgG CBA, and each row is an individual serum sample. Negative samples are black, and positive samples are red. The samples are shown according to their serostatus sent by the individual centers. (B) Agreement of assay results for IgG CBAs (assays 1–8). Sample are grouped according to their agreement in all 8 CBAs or in the 7 live CBAs (red: positive in all CBAs, black: negative in all CBAs, white: discordant). (C) Quantitative results for the 8 MOG-IgG CBA-IF and CBA-FACS assays. The cutoff values for all assays except assay Nr. 7 are indicated by the dashed gray lines. For assay Nr. 7, cutoff levels for pediatric samples (blue dots) are indicated by the blue dashed line, and cutoff levels for adult samples (red dots) are indicated by the red dashed line. The quantitative range of each assay result for its probability to be seropositive in all live CBAs is indicated by the dotted line and shaded in darker gray (100% probability), whereas the range of discordant samples is shaded in light gray. BD = blood donor; bdl neg = borderline negative values just below the individual cutoff levels; CBA = cell-based assay; FACS = fluorescence-activated cell sorting; IF = immunofluorescence; Ig = immunoglobulin; low pos = positive values just above the individual cutoff levels; MOG = myelin oligodendrocyte glycoprotein.
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