Quantitative Determination of Circulating Soluble Egg Antigen in Urine and Serum of Schistosoma mansoni-Infected Individuals Using a Combined Two-Site Enzyme-Linked Immunosorbent Assay

Mohamed Saad A. Nourel DinLaboratory of Parasitology, Medical Faculty, University of Leiden, Leiden, The Netherlands

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Ria NibbelingLaboratory of Parasitology, Medical Faculty, University of Leiden, Leiden, The Netherlands

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J. Peter RotmansLaboratory of Parasitology, Medical Faculty, University of Leiden, Leiden, The Netherlands

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Anton M. PoldermanLaboratory of Parasitology, Medical Faculty, University of Leiden, Leiden, The Netherlands

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Frederik W. KrijgerLaboratory of Parasitology, Medical Faculty, University of Leiden, Leiden, The Netherlands

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Andre M. DeelderLaboratory of Parasitology, Medical Faculty, University of Leiden, Leiden, The Netherlands

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Two enzyme-linked immunosorbent assays (ELISA-5B1 using monoclonal antibody [MAb] 114-5B1-A [IgG1] and ELISA-4D12 using MAb 114-4D12-A [IgG3]) that detect circulating soluble egg antigen (CSEA) of Schistosoma mansoni were combined into one assay. This assay showed better performance than either of the two MAbs alone in detecting egg antigen, which was demonstrated with 80 urine samples from patients infected with Schistosoma mansoni from Zaire. The lower detection limit of the combined ELISA was 90 pg of the trichloroacetic acid—soluble fraction of soluble egg antigen (SEA-TCA) per milliliter. Thirty-two serum samples and 107 urine samples from uninfected Dutch individuals were negative when tested with the combined ELISA. This assay showed the same sensitivity (86.3%) with patients' urine samples as parallel testing with ELISA-5B1 and ELISA-4D12 (85%), while ELISA-5B1 and ELISA-4D12 showed sensitivities of 81.3% and 75%, respectively. The sensitivity of the combined ELISA with 51 serum samples was 84.3%, and three of five serum samples available from the seven patients with negative urine were positive for CSEA. The concentration of CSEA calculated from a four-parameters logistic curve for samples tested showed a correlation with egg output and serum circulating anodic antigen (P < 0.0001). Circulating soluble egg antigen in urine showed a significant decrease with an increase in age of the patients in relation to serum CSEA and egg output.

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