Case Report: Diagnosis and Assessment of Cure Approaches for Acute Schistosomiasis in Pre-School Children.

Acute schistosomiasis (AS) manifests with a broad spectrum of clinical features in pediatric populations. Diagnosis may be difficult in the absence of detectable numbers of eggs. As a result, new approaches may be required to achieve an accurate diagnosis. Optimal praziquantel (PZQ) treatment regimen for young children is debatable. Also, the post-treatment response is still poorly evaluated due to the lack of reliable markers. A group of 6 children (a toddler and 5 pre-school children) and one pre-adolescent were investigated for AS clinical manifestations and followed-up for two years after treatment. Ova detection was performed by Kato-Katz (KK) and presence of Schistosoma mansoni DNA was assessed by real-time PCR (rt-PCR) in stool samples. IgG and IgE anti-Schistosoma levels and urinary antigen were detected by ELISA and point-of-care circulating cathodic antigen (POC-CCA) testing in serum and urine, respectively. AS clinical symptoms were present in 5/7 (71.4%) of the infected children, and hypereosinophilia was detected in all of them. Ova detection and serology were positive in only 3/7 (44.9%) and 4/7 (57.1%), respectively. However, real-time PCR (rt-PCR) showed the presence of Schistosoma DNA in 6/7 (85.7%) of the cases, and urinary antigen was detected in all infected children. The long-term follow-up after treatment with three doses of PZQ (80mg/kg/dose), showed high cure rates (CR) as demonstrated by the DNA-based assay as well as reduced levels of side effects. CR based on urinary antigen detection ranged from 28.6 to 100%, being the highest CR due to double testing the 2-year post-treatment samples. The results suggest that high dose and repeated treatment with PZQ might be effective for AS in young children. Also, new laboratory markers should be considered to diagnosis and monitor the drug response.

The use of the circulating cathodic antigen (CCA) urine cassette assay for the diagnosis and assessment of cure of Schistosoma mansoni infections in an endemic area of the Amazon region.

INTRODUCTION: Schistosomiasis is a poverty-related disease that affects people in 78 countries worldwide. This study aimed to evaluate the point-of-care circulating cathodic antigen (POC-CCA) test performance using sensitive parasitological methods as a reference standard (RS) in individuals before and after treatment. METHODS: The RS was established by combining the results of 16 Kato-Katz slides and the Helmintex® method. Positivity rates of the POC-CCA test and Kato-Katz and Helmintex® methods were calculated before treatment and 30 days afterward. Furthermore, the sensitivity, specificity, accuracy, and kappa coefficient before treatment were determined by comparing the methods. The cure rate was defined 30 days after treatment. RESULTS: Among the 217 participants, the RS detected a total of 63 (29.0%) positive individuals. The POC-CCA test identified 79 (36.4%) infections. The evaluation of POC-CCA test performance in relation to the RS revealed a sensitivity of 61.9%, specificity of 74.0%, accuracy of 70.5%, and kappa coefficient of 0.33. Out of the 53 remaining participants after treatment, a total of 45 (81.1%) showed egg negative results, and 8 (18.9%) were egg positive according to the RS. A total of 5 (9.4%) egg-positive and 37 (69.8%) egg-negative individuals were positive by the POC-CCA test. CONCLUSIONS: Our data show that the POC-CCA test has potential as an auxiliary tool for the diagnosis of Schistosoma mansoni infection, yielding better results than 16 Kato-Katz slides from three different stool samples. However, the immunochromatographic test lacks sufficient specificity and sensitivity for verifying the cure rate after treatment.

The use of the circulating cathodic antigen (CCA) urine cassette assay for the diagnosis and assessment of cure of Schistosoma mansoni infections in an endemic area of the Amazon region

Abstract INTRODUCTION Schistosomiasis is a poverty-related disease that affects people in 78 countries worldwide. This study aimed to evaluate the point-of-care circulating cathodic antigen (POC-CCA) test performance using sensitive parasitological methods as a reference standard (RS) in individuals before and after treatment. METHODS The RS was established by combining the results of 16 Kato-Katz slides and the Helmintex® method. Positivity rates of the POC-CCA test and Kato-Katz and Helmintex® methods were calculated before treatment and 30 days afterward. Furthermore, the sensitivity, specificity, accuracy, and kappa coefficient before treatment were determined by comparing the methods. The cure rate was defined 30 days after treatment. RESULTS Among the 217 participants, the RS detected a total of 63 (29.0%) positive individuals. The POC-CCA test identified 79 (36.4%) infections. The evaluation of POC-CCA test performance in relation to the RS revealed a sensitivity of 61.9%, specificity of 74.0%, accuracy of 70.5%, and kappa coefficient of 0.33. Out of the 53 remaining participants after treatment, a total of 45 (81.1%) showed egg negative results, and 8 (18.9%) were egg positive according to the RS. A total of 5 (9.4%) egg-positive and 37 (69.8%) egg-negative individuals were positive by the POC-CCA test. CONCLUSIONS Our data show that the POC-CCA test has potential as an auxiliary tool for the diagnosis of Schistosoma mansoni infection, yielding better results than 16 Kato-Katz slides from three different stool samples. However, the immunochromatographic test lacks sufficient specificity and sensitivity for verifying the cure rate after treatment.

Mapping Schistosoma mansoni endemicity in Rwanda: a critical assessment of geographical disparities arising from circulating cathodic antigen versus Kato-Katz diagnostics.

BACKGROUND: Schistosomiasis is a neglected tropical disease caused by Schistosoma parasites. Intervention relies on identifying high-risk regions, yet rapid Schistosoma diagnostics (Kato-Katz stool assays (KK) and circulating cathodic antigen urine assays (CCA)) yield different prevalence estimates. We mapped S. mansoni prevalence and delineated at-risk regions using a survey of schoolchildren in Rwanda, where S. mansoni is an endemic parasite. We asked if different diagnostics resulted in disparities in projected infection risk. METHODS: Infection data was obtained from a 2014 Rwandan school-based survey that used KK and CCA diagnostics. Across 386 schools screened by CCA (N = 19,217). To allow for uncertainty when interpreting ambiguous CCA trace readings, which accounted for 28.8% of total test results, we generated two presence-absence datasets: CCA trace as positive and CCA trace as negative. Samples (N = 9,175) from 185 schools were also screened by KK. We included land surface temperature (LST) and the Normalized Difference Vegetation and Normalized Difference Water Indices (NDVI, NDWI) as predictors in geostatistical regressions. FINDINGS: Across 8,647 children tested by both methods, prevalence was 35.93% for CCA trace as positive, 7.21% for CCA trace as negative and 1.95% for KK. LST was identified as a risk factor using KK, whereas NDVI was a risk factor for CCA models. Models predicted high endemicity in Northern and Western regions of Rwanda, though the CCA trace as positive model identified additional high-risk areas that were overlooked by the other methods. Estimates of current burden for children at highest risk (boys aged 5-9 years) varied by an order of magnitude, with 671,856 boys projected to be infected by CCA trace as positive and only 60,453 projected by CCA trace as negative results. CONCLUSIONS: Our findings show that people in Rwanda's Northern, Western and capital regions are at high risk of S. mansoni infection. However, variation in identification of environmental risk factors and delineation of at-risk regions using different diagnostics likely provides confusing messages to disease intervention managers. Further research and statistical analyses, such as latent class analysis, can be used to improve CCA result classification and assess its use in guiding treatment regimes.

Microarray assessment of N-glycan-specific IgE and IgG profiles associated with Schistosoma mansoni infection in rural and urban Uganda.

Core ß-1,2-xylose and α-1,3-fucose are antigenic motifs on schistosome N-glycans, as well as prominent IgE targets on some plant and insect glycoproteins. To map the association of schistosome infection with responses to these motifs, we assessed plasma IgE and IgG reactivity using microarray technology among Ugandans from rural Schistosoma mansoni (Sm)-endemic islands (n = 209), and from proximate urban communities with lower Sm exposure (n = 62). IgE and IgG responses to core ß-1,2-xylose and α-1,3-fucose modified N-glycans were higher in rural versus urban participants. Among rural participants, IgE and IgG to core ß-1,2-xylose were positively associated with Sm infection and concentration peaks coincided with the infection intensity peak in early adolescence. Responses to core α-1,3-fucose were elevated regardless of Sm infection status and peaked before the infection peak. Among urban participants, Sm infection intensity was predominantly light and positively associated with responses to both motifs. Principal component and hierarchical cluster analysis reduced the data to a set of variables that captured core ß-1,2-xylose- and α-1,3-fucose-specific responses, and confirmed associations with Sm and the rural environment. Responses to core ß-1,2-xylose and α-1,3-fucose have distinctive relationships with Sm infection and intensity that should further be explored for associations with protective immunity, and cross-reactivity with other exposures.

Mapping of Schistosoma mansoni in the Nile Delta, Egypt: Assessment of the prevalence by the circulating cathodic antigen urine assay.

In line with WHO recommendations on elimination of schistosomiasis, accurate identification of all areas of residual transmission is a key step to design and implement measures aimed at interrupting transmission in low-endemic settings. To this purpose, we assessed the prevalence of active S. mansoni infection in five pilot governorates in the Nile Delta of Egypt by examining schoolchildren (6-15 years) using the Urine-Circulating Cathodic Antigen (Urine-CCA) cassette test; we also carried out the standard Kato-Katz (KK) thick smear, the monitoring and evaluation tool employed by Egypt's national schistosomiasis control programme. Prevalence rates determined by the Urine-CCA test for all governorates were higher than those determined by KK (p<0.01). Of 35 districts surveyed in the five governorates, S. mansoni infection was detected in 19 districts (54.3%) using KK, and in 31 districts (88.6%) by Urine-CCA (χ2=9.94; P=0.0016). S. mansoni infections were detected by Urine-CCA, but not by KK in 12 districts (34.3%), and infection was not detected by either of the two diagnostic methods in four districts in Qalyubia governorate. Males and higher age-groups have significantly higher Urine-CCA prevalence rates. Based on the findings of the current S. mansoni mapping exercise, authorities of the Ministry of Health and Population (MoHP) adopted a new elimination strategy by readjusting thresholds for mass treatment with praziquantel and targeting all transmission areas. MoHP is now planning to remap in all other endemic governorates using Urine-CCA with the aim of identifying all areas of transmission where the elimination strategy should be applied.

Circulating antigen tests and urine reagent strips for diagnosis of active schistosomiasis in endemic areas.

BACKGROUND: Point-of-care (POC) tests for diagnosing schistosomiasis include tests based on circulating antigen detection and urine reagent strip tests. If they had sufficient diagnostic accuracy they could replace conventional microscopy as they provide a quicker answer and are easier to use. OBJECTIVES: To summarise the diagnostic accuracy of: a) urine reagent strip tests in detecting active Schistosoma haematobium infection, with microscopy as the reference standard; and b) circulating antigen tests for detecting active Schistosoma infection in geographical regions endemic for Schistosoma mansoni or S. haematobium or both, with microscopy as the reference standard. SEARCH METHODS: We searched the electronic databases MEDLINE, EMBASE, BIOSIS, MEDION, and Health Technology Assessment (HTA) without language restriction up to 30 June 2014. SELECTION CRITERIA: We included studies that used microscopy as the reference standard: for S. haematobium, microscopy of urine prepared by filtration, centrifugation, or sedimentation methods; and for S. mansoni, microscopy of stool by Kato-Katz thick smear. We included studies on participants residing in endemic areas only. DATA COLLECTION AND ANALYSIS: Two review authors independently extracted data, assessed quality of the data using QUADAS-2, and performed meta-analysis where appropriate. Using the variability of test thresholds, we used the hierarchical summary receiver operating characteristic (HSROC) model for all eligible tests (except the circulating cathodic antigen (CCA) POC for S. mansoni, where the bivariate random-effects model was more appropriate). We investigated heterogeneity, and carried out indirect comparisons where data were sufficient. Results for sensitivity and specificity are presented as percentages with 95% confidence intervals (CI). MAIN RESULTS: We included 90 studies; 88 from field settings in Africa. The median S. haematobium infection prevalence was 41% (range 1% to 89%) and 36% for S. mansoni (range 8% to 95%). Study design and conduct were poorly reported against current standards. Tests for S. haematobium Urine reagent test strips versus microscopyCompared to microscopy, the detection of microhaematuria on test strips had the highest sensitivity and specificity (sensitivity 75%, 95% CI 71% to 79%; specificity 87%, 95% CI 84% to 90%; 74 studies, 102,447 participants). For proteinuria, sensitivity was 61% and specificity was 82% (82,113 participants); and for leukocyturia, sensitivity was 58% and specificity 61% (1532 participants). However, the difference in overall test accuracy between the urine reagent strips for microhaematuria and proteinuria was not found to be different when we compared separate populations (P = 0.25), or when direct comparisons within the same individuals were performed (paired studies; P = 0.21).When tests were evaluated against the higher quality reference standard (when multiple samples were analysed), sensitivity was marginally lower for microhaematuria (71% vs 75%) and for proteinuria (49% vs 61%). The specificity of these tests was comparable. Antigen assayCompared to microscopy, the CCA test showed considerable heterogeneity; meta-analytic sensitivity estimate was 39%, 95% CI 6% to 73%; specificity 78%, 95% CI 55% to 100% (four studies, 901 participants). Tests for S. mansoni Compared to microscopy, the CCA test meta-analytic estimates for detecting S. mansoni at a single threshold of trace positive were: sensitivity 89% (95% CI 86% to 92%); and specificity 55% (95% CI 46% to 65%; 15 studies, 6091 participants) Against a higher quality reference standard, the sensitivity results were comparable (89% vs 88%) but specificity was higher (66% vs 55%). For the CAA test, sensitivity ranged from 47% to 94%, and specificity from 8% to 100% (4 studies, 1583 participants). AUTHORS' CONCLUSIONS: Among the evaluated tests for S. haematobium infection, microhaematuria correctly detected the largest proportions of infections and non-infections identified by microscopy.The CCA POC test for S. mansoni detects a very large proportion of infections identified by microscopy, but it misclassifies a large proportion of microscopy negatives as positives in endemic areas with a moderate to high prevalence of infection, possibly because the test is potentially more sensitive than microscopy.

Use of humanised rat basophilic leukaemia cell line RS-ATL8 for the assessment of allergenicity of Schistosoma mansoni proteins.

BACKGROUND: Parasite-specific IgE is thought to correlate with protection against Schistosoma mansoni infection or re-infection. Only a few molecular targets of the IgE response in S. mansoni infection have been characterised. A better insight into the basic mechanisms of anti-parasite immunity could be gained from a genome-wide characterisation of such S. mansoni allergens. This would have repercussions on our understanding of allergy and the development of safe and efficacious vaccinations against helminthic parasites. METHODOLOGY/PRINCIPAL FINDINGS: A complete medium- to high-throughput amenable workflow, including important quality controls, is described, which enables the rapid translation of S. mansoni proteins using wheat germ lysate and subsequent assessment of potential allergenicity with a humanised Rat Basophilic Leukemia (RBL) reporter cell line. Cell-free translation is completed within 90 minutes, generating sufficient amounts of parasitic protein for rapid screening of allergenicity without any need for purification. Antigenic integrity is demonstrated using Western Blotting. After overnight incubation with infected individuals' serum, the RS-ATL8 reporter cell line is challenged with the complete wheat germ translation mixture and Luciferase activity measured, reporting cellular activation by the suspected allergen. The suitability of this system for characterization of novel S. mansoni allergens is demonstrated using well characterised plant and parasitic allergens such as Par j 2, SmTAL-1 and the IgE binding factor IPSE/alpha-1, expressed in wheat germ lysates and/or E. coli. SmTAL-1, but not SmTAL2 (used as a negative control), was able to activate the basophil reporter cell line. CONCLUSION/SIGNIFICANCE: This method offers an accessible way for assessment of potential allergenicity of anti-helminthic vaccine candidates and is suitable for medium- to high-throughput studies using infected individual sera. It is also suitable for the study of the basis of allergenicity of helminthic proteins.

Schistosoma mansoni in a low-prevalence area in Brazil: the importance of additional methods for the diagnosis of hard-to-detect individual carriers by low-cost immunological assays.

Schistosomiasis diagnosis is based on the detection of eggs in the faeces, which is laborious and lacks sensitivity, especially for patients with a low parasite burden. Immunological assays for specific antibody detection are available, but they usually demonstrate low sensitivity and/or specificity. In this study, two simple immunological assays were evaluated for the detection of soluble Schistosoma mansoni adult worm preparation (SWAP) and egg-specific IgGs. These studies have not yet been evaluated for patients with low parasite burdens. Residents of an endemic area in Brazil donated sera and faecal samples for our study. The patients were initially diagnosed by a rigorous Kato-Katz analysis of 18 thick smears from four different stool samples. The ELISA-SWAP was successful for human diagnosis with 90% sensitivity and specificity, confirming the Kato-Katz diagnosis with nearly perfect agreement, as seen by the Kappa index (0.85). Although the ELISA-soluble S. mansoni egg antigen was 85% sensitive, it exhibited low specificity (80%; Kappa index: 0.75) and was more susceptible to cross-reactivity. We believe that immunological assays should be used in conjunction with Kato-Katz analysis as a supplementary tool for the diagnosis of schistosomiasis for patients with low infection burdens, which are usually hard to detect.

Schistosoma mansoni in a low-prevalence area in Brazil: the importance of additional methods for the diagnosis of hard-to-detect individual carriers by low-cost immunological assays

Schistosomiasis diagnosis is based on the detection of eggs in the faeces, which is laborious and lacks sensitivity, especially for patients with a low parasite burden. Immunological assays for specific antibody detection are available, but they usually demonstrate low sensitivity and/or specificity. In this study, two simple immunological assays were evaluated for the detection of soluble Schistosoma mansoni adult worm preparation (SWAP) and egg-specific IgGs. These studies have not yet been evaluated for patients with low parasite burdens. Residents of an endemic area in Brazil donated sera and faecal samples for our study. The patients were initially diagnosed by a rigorous Kato-Katz analysis of 18 thick smears from four different stool samples. The ELISA-SWAP was successful for human diagnosis with 90% sensitivity and specificity, confirming the Kato-Katz diagnosis with nearly perfect agreement, as seen by the Kappa index (0.85). Although the ELISA-soluble S. mansoni egg antigen was 85% sensitive, it exhibited low specificity (80%; Kappa index: 0.75) and was more susceptible to cross-reactivity. We believe that immunological assays should be used in conjunction with Kato-Katz analysis as a supplementary tool for the diagnosis of schistosomiasis for patients with low infection burdens, which are usually hard to detect.

Assessment of a DNA vaccine encoding an anchored-glycosylphosphatidylinositol tegumental antigen complexed to protamine sulphate on immunoprotection against murine schistosomiasis.

Protamine sulphate/DNA complexes have been shown to protect DNA from DNase digestion in a lipid system for gene transfer. A DNA-based vaccine complexed to protamine sulphate was used to induce an immune response against Schistosoma mansoni anchored-glycosylphosphatidylinositol tegumental antigen in BALB/c mice. The protection elicited ranged from 33 to 44%. The spectrum of the elicited immune response induced by the vaccine formulation without protamine was characterized by a high level of IgG (IgG1> IgG2a). Protamine sulphate added to the DNA vaccine formulation retained the green fluorescent protein encoding-plasmid longer in muscle and spleen. The experiments in vivo showed that under protamine sulphate effect, the scope of protection remained unchanged, but a modulation in antibody production (IgG1= IgG2a) was observed.

Assessment of a DNA vaccine encoding an anchored-glycosylphosphatidylinositol tegumental antigen complexed to protamine sulphate on immunoprotection against murine schistosomiasis

Protamine sulphate/DNA complexes have been shown to protect DNA from DNase digestion in a lipid system for gene transfer. A DNA-based vaccine complexed to protamine sulphate was used to induce an immune response against Schistosoma mansoni anchored-glycosylphosphatidylinositol tegumental antigen in BALB/c mice. The protection elicited ranged from 33 to 44 percent. The spectrum of the elicited immune response induced by the vaccine formulation without protamine was characterized by a high level of IgG (IgG1> IgG2a). Protamine sulphate added to the DNA vaccine formulation retained the green fluorescent protein encoding-plasmid longer in muscle and spleen. The experiments in vivo showed that under protamine sulphate effect, the scope of protection remained unchanged, but a modulation in antibody production (IgG1= IgG2a) was observed.

Practical and ethical issues in the development of a vaccine against schistosomiasis mansoni.

Clinical trials to evaluate schistosomal vaccines are in progress. We discuss the desired characteristics of such a vaccine, propose a product profile, and consider the clinical and pre-clinical studies needed for its licensure, within practical and ethical constraints. We believe that licensure of a schistosomal vaccine will be greatly facilitated by resolution of the following issues: identification of the human immunoprotective antigens and mechanisms; induction of the appropriate responses by adjuvanted vaccines; understanding the effect of immunization on immunopathology; development of an improved serologic assay to determine worm burden; generation of approximately $500 million to fund the project; and development of a physical infrastructure with trained professionals in disease-endemic countries to perform Phase III clinical trials. We also believe that development of a schistosomal vaccine, while a long range goal, is possible and desirable, and we have indicated some of the practical steps that will be required to achieve this laudable accomplishment.

Susceptibility and resistance to Schistosoma mansoni reinfection: parallel cellular and isotypic immunologic assessment.

Cellular and humoral immune responses to Schistosoma mansoni antigen preparations were evaluated in individuals presumed to be susceptible or resistant to reinfection after chemotherapeutic cure. A consistent proliferative increase in the response to soluble egg antigen (SEA) was observed post-treatment in both the susceptible and resistant groups. However, this change was not related to resistance. Isotype studies showed that IgM antibody levels to soluble worm antigen preparation (SWAP) and cercariae antigens were significantly higher in the resistant group than in the susceptible group. Post-treatment, an increase in IgE anti-SWAP and anti-schistosomular tegument (STEG) responses and a decrease in IgG4 anti-SEA and anti-STEG responses were observed in the resistant group. These finding are similar to those we have reported previously for a putative resistant group termed endemic normals, and are compatible with immunologic studies in different endemic areas. Together, these findings indicate that even on the population level, high IgE specificities coupled with low IgG4 specificities correlate well with documented resistance to reinfection.

Relating serum circulating anodic antigens to faecal egg counts in Schistosoma mansoni infections: a modelling approach.

Circulating anodic antigen (CAA) levels in serum and faecal egg counts are both quantitative measures of Schistosoma mansoni worm burdens. In this study, we have tested whether circulating anodic antigens can be included into an established egg count model. A data set with 3 repeated faecal egg count and serum CAA measurements of 50 individuals from a community in Burundi with moderate endemicity was used. By means of Monte Carlo simulation, both antigens and egg counts were related to an underlying worm pair distribution, taking into account the variation in repeated measurements (within individuals) and the variation in worm burdens (between individuals). Models with various assumptions (e.g. presence or absence of density-dependent egg production) were tested. Whereas observed and predicted egg counts agreed fairly well, the circulating antigen data could not be described satisfactorily. In particular, the predicted number of negative antigen concentrations was much lower than observed, while the number of light positives was overestimated. There seems to be a mechanism that causes a shift of expected (low) positive CAA concentrations towards zeros, which the proposed models do not provide for. Possible biological as well as assay-related mechanisms that may account for this shift are discussed. The assumption that serum CAA concentrations are a simple direct reflection of worm (pair) burdens could not be corroborated by this modelling exercise. Apparently, the relationship between (measured) CAA concentrations, egg counts and worm burdens in human S. mansoni infections is more complex than assumed.

Schistosoma mansoni egg specific antibodies and circulating antigens: assessment of their validity in immunodiagnosis of schistosomiasis.

A total of 127 individuals of different age and sex; 92 from Kafr-Sendewa, Qualyobia Governorate, Egypt, in comparison to 23 cases with hydatidosis and fascioliasis as a parasitic control group, and 12 healthy control group from non-endemic area. All cases were screened by clinical examination, urine, stool, rectal snip, abdominal ultrasonographic examination and indirect haemagglutination test (IHAT). Accordingly, they were grouped into active intestinal schistosomiasis group, seropositive group by (IHAT), normal control group from the same endemic area, parasitic control group and normal control group. All cases were subjected to detection of IgG, IgM, IgG4, anti-soluble egg antigen (SEA) and anti-excretory-secretory egg antigen (ESEA) by ELISA tests; and circulating egg antigens by double-sandwich ELISA techniques. The results showed that IgG4 anti-SEA is the best diagnostic test, as it gave the best diagnostic efficacy (90%). Also, it is a good screening test which can be used in endemic area as it gave significant difference between the active intestinal schistosomiasis cases with each of the endemic control group (P < 0.001) and the seropositive cases (P < 0.05). Other valid diagnostic egg specific antibodies tests were IgM anti-SEA and IgG anti-SEA as their diagnostic efficacy were 80% and 76.6% respectively (P < 0.05). The diagnostic efficacy of circulating antigen detection (C.Ag) test was 70% (P < 0.05). In addition, it was the most specific test with 100% specificity. IG4-anti-ESEA gave the least cross-reaction with other parasites (17.3%). The mean optical density (OD) level of circulating antigen detection test was significantly higher in the organomegalic (hepatosplenomegaly) cases than the non-organomegalic cases (P < 0.05).

Dificuldades no desenvolvimento de uma vacina para esquistossomose mansoni / Problems in the development of a vaccine against shistosomiasis mansoni

Nas últimas duas décadas, os estudos com antígenos que possam conferir imunidade protetora à infecçäo experimental pelo Schistosoma mansoni tiveram grande impulso, especialmente devido ao avanço dos conhecimentos nos campos da biologia molecular e da imunologia. Embora, estes estudos tenham trazidos novas e importantes contribuiçöes, até o momento, a proteçäo conferida tem girado em torno de 50 por cento de diminuiçäo do número de vermes e com alguns antígenos, também do número de ovos. As muitas perguntas ainda näo respondidas indicam a necessidade de novas pesquisas a serem realizadas em animais de pequeno e grande porte, antes que estes antígenos candidatos a vacina, possam ser utilizados em ensaios clínicos

Seroepidemiology of schistosomiasis in Puerto Rico: evidence for vanishing endemicity.

The current study summarizes our findings of anti-schistosome egg antibody by the circumoval precipitin test for two different populations in Puerto Rico. One group, exclusively males more than 40 years of age and from all municipalities on the island, was from the Veterans Administration Hospital for the period 1988-1997. The second group resided southeast of San Juan, around the municipality of Caguas and adjacent municipalities east of Caguas, was of both sexes and mostly until 1997 of undetermined ages for the period 1993-1997. Results reveal a yearly decrease in testing requests from the Veterans Administration Hospital from 148 in 1988 and 1989 with 16% positive to three in 1996 through 1998 with none positive. This decrease in testing requests was because of a decrease of suspicion of schistosomiasis in this group. The other patient population from the Caguas region showed a gradual but continuous decrease in seropositive individuals from 21% in 1993 to 12% in 1996, with precipitous decrease to 5% in 1997 and only 1% in 1998. Moreover, there were four patients from which at least two serum samples were obtained one or two years apart and tested. In each instance the more recently obtained sample had lower antibody reactions than the first as reflected in lower percentages of positive egg reactors. The fact that they were treated with praziquantel after the first testing also suggests that the infected population was being eliminated through chemotherapy. These combined results suggest the elimination of infections with Schistosoma mansoni in the traditionally high prevalence regions east of San Juan in the absence of any proactive control efforts in Puerto Rico. Because of the rapid urbanizing of Puerto Rico, the one identifiable control effort is economic development and well being.

Circulating schistosomal antigen in diagnosis and assessment of cure in individuals infected with Schistosoma mansoni.

The effectiveness of praziquantel in treating schistosomiasis is most commonly assessed by quantitating egg production or anti-schistosome antibodies in serum. We have used a monoclonal antibody (MAb)-based antigen-capture enzyme-linked immunosorbent assay (ELISA) for the serologic diagnosis of schistosomiasis, and to monitor the efficacy of praziquantel therapy in 49 individuals with parasitologically proven schistosomiasis. The MAb used, 128C3/3/21, recognizes a repeating carbohydrate epitope expressed at all stages of parasite development, and antibodies recognizing this epitope are found in the serum of infected humans. The overall sensitivity of the ELISA was 78%, with a sensitivity of 100% for patients excreting greater than 100 eggs/g of feces and 72% for those excreting less than 100 eggs/g of feces. The positivity of the ELISA was directly related to the fecal egg counts obtained on days -3, -2, and -1 before treatment with praziquantel, but there was no correlation between antigen levels and the clinical stage of the disease. After praziquantel treatment, we observed a highly significant correlation (P less than 0.0001) between the time elapsed since treatment and the decrease in antigenemia. Furthermore, although no eggs were detected in any of the stool specimens at week 12 after treatment, the antigen was detected in 21% of the treated patients (seven of 33 ELISA-positive patients). Antigen levels decreased over the 12-week period in six of these patients, whereas the antigen level increased with time in one individual. The persistence of antigenemia suggests that these individuals are either still clearing antigen or remain infected.(ABSTRACT TRUNCATED AT 250 WORDS)

Assessment of cure in schistosomiasis patients after chemotherapy with praziquantel by quantitation of circulating anodic antigen (CAA) in urine.

The kinetics of circulating anodic antigen (CAA) levels in urine were studied in Egyptian male patients infected with Schistosoma mansoni or with both S. mansoni and S. haematobium, before treatment, and at one, three and six weeks after chemotherapy. A quantitative enzyme-linked immunosorbent assay (ELISA) demonstrated CAA in 82% of the serum and 89% of the urine samples from these 28 patients. To evaluate the possibility of circadian variability in urine CAA levels, samples were examined in 15 patients at four intervals during a 24-hour period. No significant differences in CAA titers were observed. Seventeen patients were subsequently treated with praziquantel and followed for six weeks. CAA titers in serum and urine decreased significantly one week after therapy. Thereafter, the profile of CAA titer in urine continued to show a parallel but delayed decline compared to that in serum. While all serum CAA titers became negative three to six weeks after treatment, urine titers were negative in 47% at three weeks and 69% at six weeks. The remaining positive patients had low titers. A significant quantitative correlation in CAA titer was found between serum and urine before and after treatment. Seventeen Egyptian control subjects with no active schistosome infection were negative for CAA in both serum and urine. Our results confirm that the CAA urine assay could be used as a sensitive and non-invasive method to diagnose the disease, and indicate that the assay can be used to monitor efficacy of schistosome chemotherapy.