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Fig 1.

Inclusion and exclusion criteria of the study participants.

Of the 400 pupils enrolled in the three schools, 269 were included in the analysis.

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Table 1.

S. haematobium infection prevalence and intensity determined by the results of pCAA500 and egg detection among 269 pupils.

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Fig 2.

Individuals with active S. haematobium infection showed higher IgG levels against the four antigen sets.

Total IgG levels against (a) ShSEA, (b) ShSerpin, (c) RP26, (d) ShSerpin-RP26 mixture were analyzed among individuals from Kwale with active S. haematobium infection (Sh+, n = 141, shown in red dots) and without active infection (Sh-, n = 128, blue dots) determined by the composite reference (CAA Indecisive results regarded as CAA negative, Indec-). The non-endemic control samples are included as the reference (Non endemic, n = 25, black dots). The dotted lines show the cut-off values determined by the geometric mean plus 3 SD of non-endemic controls (Japanese, n = 25)’ unit values. The cut-off value of each antigen was; 0.459 for ShSEA, 0.552 for ShSerpin, 0.165 for RP26, and 0.155 for ShSerpin-RP26 mixture. The antibody levels of the Sh+ and Sh- groups were compared by using Mann-Whitney’s U tests. Statistical significance was set at p < 0.05 and is shown using asterisks: **** = p < 0.0001. The horizontal bars represent the median values of arbitrary units of each group. Sh+: S. haematobium infection positive (pCAA500 positive and/or urine egg positive), shown in red dots (n = 141). Sh-: S. haematobium infection negative (pCAA500 negative and urine egg negative), shown in blue dots (n = 128). NC: non-endemic controls (Japanese), shown in black dots (n = 25).

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Fig 3.

The ShSerpin-RP26 mixture capacity detecting active infections was similar to ShSEA.

The ROC curves were generated from the ELISA results of Kwale samples infected (Sh+, n = 141) and uninfected (Sh-, n = 128) with S. haematobium. pCAA500 indecisive results were considered as CAA negative (Inde-). The area under curve (AUC) of ShSEA, ShSerpin, RP26 and ShSerpin-RP26 mixture was 0.884, 0.819, 0.792 and 0.887, respectively.

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Table 2.

Diagnostic performance of total IgG detection against ShSEA, ShSerpin, RP26 and ShSerpin-RP26 mix.

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Fig 4.

IgG levels against S. haematobium antigens were associated with the infection intensity by plasma CAA.

Total IgG levels against (a) ShSEA, (b) ShSerpin, (c) RP26, (d) ShSerpin-RP26 mixture were analyzed among Kwale samples with different categories of infection intensity determined by plasma CAA concentrations (pCAA500). CAA High: pCAA500 ≥ 100 pg/mL (n = 42); CAA Medium: 10 ≤ pCAA500 < 100 pg/mL (n = 41); CAA Low:1 ≤ pCAA500 < 10 pg/mL (n = 52); CAA Indecisive: 0.5 ≤ pCAA500 < 1 pg/mL (n = 20); CAA Negative: 0 ≤ pCAA500 < 0.5 pg/mL (n = 114). Kruskal-Wallis tests were performed for comparing antibody levels between different infection intensity groups measured by PCAA. Statistical significance was set at p < 0.05 and is shown using asterisks: **** = p < 0.0001. The dotted lines show the cut-off values determined by the geometric mean plus 3 SD of non-endemic controls’ unit values. The cut-off value of each antigen was; 0.459 for ShSEA, 0.552 for ShSerpin, 0.165 for RP26, and 0.155 for ShSerpin-RP26 mixture. The horizontal bars represent the median values of arbitrary units of each group.

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Fig 5.

Correlation between the CAA concentration by pCAA500 and the total IgG levels against ShSEA, ShSerpin, RP26 and ShSerpin-RP26 mixture. The correlations between total IgG levels against (a) ShSEA, (b) ShSerpin, (c) RP26, (d) ShSerpin-RP26 mixture and CAA concentrations were analyzed using Spearman’s rank correlation coefficient analyses. The correlation coefficient of ShSEA, ShSerpin, RP26 and ShSerpin-RP26 mixture was 0.729 (95% CI 0.665–0.782), 0.630 (95% CI 0.550–0.699), 0.540 (95% CI 0.447–0.622) and 0.695 (95% CI 0.625–0.754) respectively. For the all antigen sets, the p values were below 0.0001. As the axes are logarithmic, actual CAA concentration + 1 were plotted in the figures.

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Fig 6.

Recombinant protein antigens (ShSerpin, RP26 and ShSerpin-RP26 mixture) had lower crosss-reactivity compared to ShSEA.

Total IgG levels against (a) ShSEA, (b) ShSerpin, (c) RP26, (d) ShSerpin-RP26 mix were analyzed among 131 samples with helminthic infections. The samples were from patients with schistosomiasis mansoni (Sm, n = 20), schistosomiasis japonica (Sj, n = 12), paragonimiasis (Pw, n = 20), fascioliasis (Fh, n = 20), clonorchiasis (Cs, n = 10), sparganosis (Se, n = 19), gnathostomiasis (Gd, n = 10) and toxocariasis (Tc, n = 20). The dotted lines show the cut-off values determined by the geometric mean plus 3 SD of non-endemic controls’ unit values. The cut-off value of each antigen was; 0.459 for ShSEA, 0.552 for ShSerpin, 0.165 for RP26, and 0.155 for ShSerpin-RP26 mixture. The horizontal bars represent the median values of arbitrary units of each type of parasite infection.

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Table 3.

Number and proportion of cross-reactive samples from different helminthic infections.

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