Steere AC, Jameson JL, Fauci AS, Kasper DL, Hauser SL, Longo DL, Loscalzo J. and plate incubation procedures (5). The string mV indicates linkage to a altered short sequence of the C6 peptide (15, 16). Biotinylated antigen was purchased from Bio-Rad (Hercules, CA, USA) and bound to streptavidin-coated beads. Pooled beads were incubated with serum followed by anti-human IgG-phycoerythrin. Mean fluorescence intensity (MFI) minus fluorescence background was quantified (5). Selection of potentially useful antigens. Thrombin Inhibitor 2 Antigens were excluded from further analysis if no meaningful overall variance was detected (standard error of MFI?of 50 over all samples; comparisons, (i) age of patient and (ii) disease stage (early versus early disseminated versus late Lyme disease). For age of patient, reactivity was assessed by Pearsons (unadjusted values), and patients were divided into three age Thrombin Inhibitor 2 groups: 1 to 7, 8 to 12, and 13 to 21?years. An expression score was calculated as the MFI of the 27-antigen panel. Distribution differences in the expression scores of different age ranges were assessed by a 3-sample Anderson-Darling test. For disease stage, expression Thrombin Inhibitor 2 was separated by stage, and the Kruskal-Wallis test was then used per antigen, followed by Dunns test for the pairwise comparisons; values were adjusted by Holms correction. was used as an estimate of effect size for the Kruskal-Wallis test: (statistic) (17). Stage dependency was further assessed by principal component (PC) analysis with reactivity against the 27 antigens calculated and plotted in both 2 and 3 dimensions. Quality of representation (cos2) was calculated for the 2-dimension scenario and compared with the 3-dimension scenario. Intercorrelation of variables with PC1 and PC2 was visualized in a correlation circle, and the relative contribution of variables to each PC was calculated as cos2 divided by the total cos2 of each PC. Contributions Thrombin Inhibitor 2 of antigens to the first three PCs were compared with a hypothetical uniform variable contribution scenario. Nonnormality was confirmed by Shapiro-Wilk. We then used the Mann-Whitney U test with value adjustment by Holms correction, followed by a calculation of (Z, Z statistic; statistic (= sensitivity + specificity C 1) was employed as the optimal cut point for the estimated probability to maximize both sensitivity and specificity. Sensitivity and specificity were reported around the cohort that received valid results in the antigens that were not dropped by the regression. Confidence intervals were calculated with 2,000 stratified bootstrap replicates. We used Fishers exact test to compare proportions. We used R (v4.0.4) and BioRender.com for statistical analyses and visualization (19,C28). RESULTS Patient population. We enrolled 583 children across the 4 Pedi Lyme Net sites between June 2015 and October 2016, of whom 527 (90%) met initial study inclusion criteria (Table 1). Overall, 127 (24%) had Lyme disease and 400 (76%) were clinical mimics. Among the 127 children with Lyme disease, 13 had a single EM lesion and were considered early Lyme, whereas 55 had early disseminated and 59 late Lyme disease. TABLE 1 Clinical characteristics of enrolled children with and without Lyme disease ?[no./total no. (%)]103/127 (81)320/400 (80)Early (single EM lesion) [no./total no. (%)]13/127 (10)NAantigens Rabbit Polyclonal to 53BP1 to discriminate Lyme patients from clinical mimic. Uninformative antigens (Fig. S1) were filtered as stated in Materials and Methods. Asterisks indicate significance. Early diss., early disseminated. Distribution of antigen expression as a function of case definition. For significant comparisons (is usually shown as a measure of effect size. Asterisks indicate statistical significance according to the respective value (*, 0.05, **, 0.01; ***, 0.001; ****, 0.0001). Serologic reactivity to antigens does not differ by age. To assess whether patient age correlated with antigen.
Steere AC, Jameson JL, Fauci AS, Kasper DL, Hauser SL, Longo DL, Loscalzo J