In this study we describe the ex cretion patterns of circulating anodic (CAA) and circulating cathodic antigen (CCA) by freshly transformed and developing Schistosoma mansoni schistosomula and by adult worms, in vitro and in vivo. In vitro, CA A and CCA were detected in the culture med iu m of the para sites manuscript submitted Congress for Tropical Medicine an d Malaria

We have developed an enzyme immunoassay (ELISA) for the quantification of the schistosome circulating cathodic antigen (CCA), a glycoprotein associated with the syncitium lining the gut of the parasite. A mouse monoclonal antibody of IgG3 isotype was used as coating (antigen-capture) antibody, while a biotinylated mouse monoclonal IgM was used as second (antigen-detecting) antibody. Streptavidin-alkaline phosphatase was used as enzyme label. The lower detection limit of the assay was 1.0 ng of the trichloroacetic acid soluble fraction of adult worm antigen (AWA-TCA) per ml, which corresponds to approximately 0.2 ng CCA per ml. The ELISA showed a linear range from 1.0 to 62.5 ng AWA-TCA per ml. Serum and urine samples of 16 individuals infected with Schistosoma mansoni (egg counts ranging from 5 to 4820 eggs per gram of faeces) were tested in the assay. Antigen titres ranged from less than 4-8192. This assay represents a considerable advantage in diagnosis of Schistosoma infections as it allows the detection and quantification of CCA in serum and urine in even lightly infected individuals.

Using spleen cells of mice infected or immunized respectively with cercariae or antigen preparations of Schistosoma mansoni, S. haematobium or S. japonicum monoclonal antibodies (mAbs) were produced against the schistosome gut-associated antigens CAA (circulating anodic antigen) and CCA (circulating cathodic antigen). Fusions nearly exclusively produced either anti-CAA (n = 25) or anti-CCA mAbs (n = 55) with a strong isotype restriction (IgM, IgG1 and IgG3) against both antigens, the majority of anti-CAA mAbs being IgG1 and the majority of anti-CCA mAbs being IgM. The mAbs, which on the basis of their selection were reactive with multiple carbohydrate epitopes of CAA or CCA, were applied in different immunological techniques including immunofluorescence, a dot immunobinding assay and immunoelectrophoresis to study the epitope repertoire. Anti-CAA mAbs were found to be reactive with 5 different epitopes, none of which occurred as multiple epitopes on eggs. Anti-CCA mAbs, on the other hand, recognized at least 10 different epitopes, while 44% of anti-CCA mAbs recognized epitopes common to the adult worm and the egg. Both CAA- and CCA-epitopes were found to be developmentally expressed at the level of the tegument in cercariae, schistosomula and 5-day-old lung worms, but in the adult worm were primarily found in the gut. Thus, the production of panels of mAbs has not only resulted in the selection of reagents optimally performing in diagnostic immunoassays, but also allowed a more detailed study of the epitope repertoire of these important schistosome antigens.

A panel of 17 monoclonal antibodies (MAbs) against Schistosoma soluble egg antigens (SEAs) was produced from BALB/c mice immunized with antigens secreted/excreted by Schistosoma mansoni eggs. In this study, we demonstrate that 16 MAbs were reactive with S. haematobium SEA in addition to S. mansoni SEA. The MAbs were tested as potential immunodiagnostic reagents in a homologous sandwich ELISA format to detect circulating soluble egg antigens (CSEAs) in serum and urine samples of S. mansoni-or S. haematobium-infected individuals. When samples of S. mansoni-infected individuals were tested, none of these MAbs performed as good as the previously described S. mansoni-specific 114-5B1-A and 114-4D12-A MAbs. However, 11 MAbs (of the IgM isotype) detected CSEA in urine samples of S. haematobium-infected individuals. Three MAbs, 290-2E6-A, 291-3D5-A, and 291-5D5-A, were selected for a pilot study with 47 urine samples of S. haematobium-infected individuals from Kenya. The CSEA levels detected with each of these ELISAs showed a significant correlation with urinary egg counts (Spearman rho Ͼ 0.37, P Ͻ 0.01) and with each other (Spearman rho Ͼ 0.74, P Ͻ 0.001). Based on the 92% specificity and 90% sensitivity of the assay, the ELISA using MAb 290-2E6-A was found to be the most promising assay for immunodiagnosis of S. haematobium infections.

Abstract. While research on alternative diagnostic and morbidity markers for infection with Schistosoma haematobium has been going on for a long time, egg counts continue to be used as the gold standard, and infection intensity is thought to reflect the severity of the disease. However, this relationship is not always clear and fluctuation in egg output makes it difficult to classify prevalence correctly. The use of circulating adult worm antigen detection as an alternative diagnostic technique has been applied with varying success. However, this is a measure of worm burden and does not reflect the tissue egg load(s). In the present study we have used an assay that detects soluble egg antigen (SEA) in urine of S. haematobium-infected children, and we have evaluated the applicability of the assay as a diagnostic and morbidity indicator. To evaluate this assay, we have studied a group of 470 children from two schools (Tsunguni and Kibaokiche) in the Coast province of Kenya; 84.8% and 77% were egg-positive while the percentage positive as determined by the SEA-ELISA were 78.8% and 76.2% in Tsunguni and Kibaokiche, respectively. In both schools, SEA levels in urine of S. haematobium-infected children significantly correlated with egg counts (Pearson's r ϭ 0.73, P Ͻ 0.0001) and with hematuria (Spearman's r ϭ 0.65, P Ͻ 0.0001). In addition, urinary tract pathology as determined by ultrasound significantly correlated with the SEA levels in urine (Spearman's r ϭ 0.3, P Ͻ 0.001). The SEA-ELISA compared well with microhematuria within egg count classes and with egg counts within hematuria classes.Detection of Schistosoma haematobium ova in urine of infected individuals remains the leading method for the direct diagnosis of the disease, and the intensity of infection expressed as number of eggs per 10 ml of urine is thought to reflect the worm burden. However, a homogeneous distribution of S. haematobium ova in urine is difficult to achieve, 1 while day-to-day and circadian variation in egg excretion may lead to incorrect estimates in prevalence and intensity of infection. Additionally, the relationship between egg counts and pathology is not always clear. 2 The presence and intensity of microhematuria, proteinuria, and leukocyturia are semi-quantitative measures of S. haematobium infection, and these parameters were found to correlate with infection intensity. 3-5 However, variations dependent on geographic area, endemicity, cultural practices, age, sex, and even time of the day exist, limiting the application of hematuria as a diagnostic tool. 6-9 Ultrasound scanning of the urinary tract remains the gold standard for pathology. This technique has been applied in the field and correlates well with standard direct and indirect measurements of S. haematobium morbidity. 10,11 However, ultrasound equipment is expensive and requires trained physicians to handle the equipment and interpret the results, limiting its use under field conditions. These shortcomings have led to the search for alternative quantitative diagnostic meth...