Detection Of Circulating Antigen Research Articles

Diagnosis of Schistosomiasis without a Microscope: Evaluating Circulating Antigen (CCA, CAA) and DNA Detection Methods on Banked Samples of a Community-Based Survey from DR Congo.

Detection of Schistosoma eggs in stool or urine is known for its low sensitivity in diagnosing light infections. Alternative diagnostics with better sensitivity while remaining highly specific, such as real-time PCR and circulating antigen detection, are progressively used as complementary diagnostic procedures but have not yet replaced microscopy. This study evaluates these alternative methods for the detection of Schistosoma infections in the absence of microscopy. Schistosomiasis presence was determined retrospectively in 314 banked stool and urine samples, available from a previous survey on the prevalence of taeniasis in a community in the Democratic Republic of the Congo, using real-time PCR, the point-of-care circulating cathodic antigen (POC-CCA) test, as well as the up-converting particle lateral flow circulating anodic antigen (UCP-LF CAA) test. Schistosoma DNA was present in urine (3%) and stool (28%) samples, while CCA (28%) and CAA (69%) were detected in urine. Further analysis of the generated data indicated stool-based PCR and the POC-CCA test to be suitable diagnostics for screening of S. mansoni infections, even in the absence of microscopy. A substantial proportion (60%) of the 215 CAA-positive cases showed low antigen concentrations, suggesting that even PCR and POC-CCA underestimated the “true” number of schistosome positives.

Re-visiting the detection of porcine cysticercosis based on full carcass dissections of naturally Taenia solium infected pigs

BackgroundTaenia solium is a neglected zoonotic parasite. The performances of existing tools for the diagnosis of porcine cysticercosis need further assessment, and their shortcomings call for alternatives. The objective of this study was to evaluate the performance of tongue palpation and circulating antigen detection for the detection of porcine cysticercosis in naturally infected pigs of slaughter age compared to full carcass dissections (considered the gold standard). Additionally, alternative postmortem dissection procedures were investigated. A total of 68 rural pigs of slaughter age randomly selected in the Eastern Province of Zambia were dissected. Dissections were conducted on full carcasses (or half carcass in case cysticerci were already detected in the first half), including all the organs. Total cysticercus counts, location and stages were recorded and collected cysticerci were identified morphologically and molecularly. All sera were analysed with the B158/B60 antigen detecting ELISA (Ag-ELISA).ResultsKey findings were the high occurrence of T. solium infected pigs (56%) and the presence of T. solium cysticerci in the livers of 26% of infected animals. More than half of the infected carcasses contained viable cysticerci. Seven carcasses had T. hydatigena cysticerci (10%), out of which five carcasses were co-infected with T. hydatigena and T. solium; two carcasses (3%) had only T. hydatigena cysticerci. Compared to full carcass dissection, the specificity of the Ag-ELISA to detect infected carcasses was estimated at 67%, the sensitivity at 68%, increasing to 90% and 100% for the detection of carcasses with one or more viable cysticerci, and more than 10 viable cysts, respectively. Tongue palpation only detected 10% of the cases, half carcass dissection 84%.Selective dissection of the diaphragm, tongue and heart or masseters can be considered, with an estimated sensitivity of 71%, increasing to 86% in carcasses with more than 10 cysticerci.ConclusionsDepending on the aim of the diagnosis, a combination of Ag-ELISA and selective dissection, including investigating the presence of T. hydatigena, can be considered. Full carcass dissection should include the dissection of the liver, kidneys, spleen and lungs, and results should be interpreted carefully, as small cysticerci can easily be overlooked.

Comparison of novel and standard diagnostic tools for the detection of Schistosoma mekongi infection in Lao People\u2019s Democratic Republic and Cambodia

BackgroundGiven the restricted distribution of Schistosoma mekongi in one province in Lao People’s Democratic Republic (Lao PDR) and two provinces in Cambodia, together with progress of the national control programmes aimed at reducing morbidity and infection prevalence, the elimination of schistosomiasis mekongi seems feasible. However, sensitive diagnostic tools will be required to determine whether elimination has been achieved. We compared several standard and novel diagnostic tools in S. mekongi-endemic areas.MethodsThe prevalence and infection intensity of S. mekongi were evaluated in 377 study participants from four villages in the endemic areas in Lao PDR and Cambodia using Kato-Katz stool examination, antibody detection based on an enzyme-linked immunosorbent assay (ELISA) and schistosome circulating antigen detection by lateral-flow tests. Two highly sensitive test systems for the detection of cathodic and anodic circulating antigens (CCA, CAA) in urine and serum were utilized.ResultsStool microscopy revealed an overall prevalence of S. mekongi of 6.4% (one case in Cambodia and 23 cases in Lao PDR), while that of Opisthorchis viverrini, hookworm, Trichuris trichiura, Ascaris lumbricoides and Taenia spp. were 50.4%, 28.1%, 3.5%, 0.3% and 1.9%, respectively. In the urine samples, the tests for CCA and CAA detected S. mekongi infections in 21.0% and 38.7% of the study participants, respectively. In the serum samples, the CAA assay revealed a prevalence of 32.4%, while a combination of the CAA assay in serum and in urine revealed a prevalence of 43.2%. There was a difference between the two study locations with a higher prevalence reached in the samples from Lao PDR.ConclusionsThe CCA, CAA and ELISA results showed substantially higher prevalence estimates for S. mekongi compared to Kato-Katz thick smears. Active schistosomiasis mekongi in Lao PDR and Cambodia might thus have been considerably underestimated previously. Hence, sustained control efforts are still needed to break transmission of S. mekongi. The pivotal role of highly sensitive diagnostic assays in areas targeting elimination cannot be overemphasised.

Wuchereria bancrofti microfilariae and quantitative circulating antigen detection in selected endemic areas in Egypt

Back ground and objectiveWuchereria bancrofti is responsible for 90% of cases of lymphatic filariasis throughout the tropics and in some subtropical areas worldwide, including Egypt. To combat this disease, the WHO has launched a program aiming to eliminate lymphatic filariasis by the year 2020 in all the endemic countries using mass drug administration (MDA) to interrupt the disease's transmission. The aim of the present work was to study W. bancrofti infection in selected endemic areas in Egypt by performing parasitological examination and enzyme-linked immunosorbent assay (ELISA) antigen detection test, and to analyze the demographic, clinical, and MDA data of the study population in relation to W. bancrofti infection.Patients and methodsA total of 300 blood samples were collected from residents in endemic areas in five governorates. Parasitological examination and Og4C3 ELISA test were performed to identify W. bancrofti infection.ResultsMicrofilariae were identified in one individual while circulating filarial antigens (CFAs) were detected in 10 individuals. Statistical analysis of the collected data showed that CFAs were significantly higher in the male population than in the female population, whereas analysis regarding other demographic, clinical, and MDA data showed no statistical significance.ConclusionThe study results showed that CFAs are still detected in endemic communities in Egypt, and that the prevalence is higher in the male population than in the female population. Although the Og4C3 ELISA test is a useful research tool for the study of W. bancrofti infections, its cost and format hinder its wide use in endemic areas.

Wuchereria bancrofti microfilariae and quantitative circulating antigen detection in selected endemic areas in Egypt

Back ground and objective Wuchereria bancrofti is responsible for 90% of cases of lymphatic filariasis throughout the tropics and in some subtropical areas worldwide, including Egypt. To combat this disease, the WHO has launched a program aiming to eliminate lymphatic filariasis by the year 2020 in all the endemic countries using mass drug administration (MDA) to interrupt the disease's transmission. The aim of the present work was to study W. bancrofti infection in selected endemic areas in Egypt by performing parasitological examination and enzyme-linked immunosorbent assay (ELISA) antigen detection test, and to analyze the demographic, clinical, and MDA data of the study population in relation to W. bancrofti infection. Patients and methods A total of 300 blood samples were collected from residents in endemic areas in five governorates. Parasitological examination and Og4C3 ELISA test were performed to identify W. bancrofti infection. Results Microfilariae were identified in one individual while circulating filarial antigens (CFAs) were detected in 10 individuals. Statistical analysis of the collected data showed that CFAs were significantly higher in the male population than in the female population, whereas analysis regarding other demographic, clinical, and MDA data showed no statistical significance. Conclusion The study results showed that CFAs are still detected in endemic communities in Egypt, and that the prevalence is higher in the male population than in the female population. Although the Og4C3 ELISA test is a useful research tool for the study of W. bancrofti infections, its cost and format hinder its wide use in endemic areas.

Taenia solium Human Cysticercosis: A Systematic Review of Sero-epidemiological Data from Endemic Zones around the World.

Background Taenia solium cysticercosis is a zoonotic neglected disease responsible for severe health disorders such as seizures and death. Understanding the epidemiology of human cysticercosis (HCC) in endemic regions will help to expose critical information about the transmission of the disease, which could be used to design efficient control programs. This review gathered serological data on apparent prevalence of T. solium circulating antigens and/or seroprevalence of T. solium antibodies, apparent prevalence of human taeniasis and risk factors for HCC from endemic communities in order to understand the differences in exposure to the parasite and active infections with T. solium metacestodes in endemic areas around the world.MethodsThree databases were used to search sero-epidemiological data from community-based studies conducted between 1989 and 2014 in cysticercosis endemic communities worldwide. The search focused on data obtained from T. solium circulating antigen detection by monoclonal antibody-based sandwich ELISA and/or T. solium antibody seroprevalence determined by Enzyme-linked Immunoelectrotransfer Blot (EITB). A meta-analysis was performed per continent.Principal FindingsA total of 39,271 participants from 19 countries, described in 37 articles were studied. The estimates for the prevalence of circulating T. solium antigens for Africa, Latin America and Asia were: 7.30% (95% CI [4.23–12.31]), 4.08% (95% CI [2.77–5.95]) and 3.98% (95% CI [2.81–5.61]), respectively. Seroprevalence estimates of T. solium antibodies were 17.37% (95% CI [3.33–56.20]), 13.03% (95% CI [9.95–16.88]) and 15.68% (95% CI [10.25–23.24]) respectively. Taeniasis reported prevalences ranged from 0 (95% CI [0.00–1.62]) to 17.25% (95% CI [14.55–20.23]).SignificanceA significant variation in the sero-epidemiological data was observed within each continent, with African countries reporting the highest apparent prevalences of active infections. Intrinsic factors in the human host such as age and immunity were main determinants for the occurrence of infections, while exposure was mostly related to environmental factors which varied from community to community.

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

BackgroundPoint‐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.ObjectivesTo 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 methodsWe searched the electronic databases MEDLINE, EMBASE, BIOSIS, MEDION, and Health Technology Assessment (HTA) without language restriction up to 30 June 2014.Selection criteriaWe 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 analysisTwo 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 resultsWe 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. haematobiumUrine 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. mansoniCompared 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% (four studies, 1583 participants).Authors' conclusionsAmong 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.23 April 2019No update plannedOtherReliable evidence with clear conclusions. All eligible published studies found in the last search (30 Jun, 2014) were included.

Preliminary evaluation of Community-Led Total Sanitation for the control of Taenia solium cysticercosis in Katete District of Zambia

Taenia solium taeniasis/cysticercosis is a zoonotic disease endemic in sub-Saharan Africa. It is associated with poor sanitary practices, free-range pig husbandry and lack of disease awareness in endemic communities. A comparative research was conducted with pre and post-intervention assessments in nine villages to evaluate Community-Led Total Sanitation (CLTS) as an intervention measure for the control of porcine cysticercosis in Katete District in the Eastern Province of Zambia. Blood samples were collected from pigs for circulating antigen detection and a questionnaire focused on the household was administered to a total of 153 respondents whose pigs were examined (64 pre-intervention, 89 post-intervention), in order to obtain information on general demographic characteristics, pig husbandry practices, sanitation practices and associated knowledge and awareness of T. solium infections. The first sampling was conducted prior to the implementation of the CLTS and second sampling eight months after triggering of CLTS in the selected villages. A total of 379 pig serum samples were examined using the B158/B60 Ag-ELISA to detect T. solium cysticercosis, 104 pre-intervention and 275 post-intervention, of which 14 (13.5%) and 45 (16.4%) were positive, respectively. Wald test p-values were computed to assess significant differences in the variables of interest mentioned above for the pre and post CLTS. The research revealed that CLTS as a control measure did not significantly improve T. solium infections in pigs. The research also revealed that the sanitation practices and awareness of cysticercosis did not change. It is recommended that a longer term evaluation be undertaken when the villages have been declared open defaecation free. In addition, the research recommends that health education, mass drug treatment and pig vaccination be incorporated, as an essential component of prevention and control programmes for T. solium infections.

Using specific synthetic peptide (p176) derived AgB 8/1-kDa accompanied by modified patient’s sera: A novel hypothesis to follow-up of Cystic echinococcosis after surgery

Cystic echinococcosis (CE) is caused by the larval stage of the tapeworm Echinococcus granulosus. Until now, CE does not have an effective follow-up after surgery. To date, CE follow-up is conducted based on either antibody or antigen detection assays by double antibody sandwich ELISA. Unlike antigen detection, antibodies to imply exposure to an Echinococcus infection while their titration could remain for a longer period (1–10years) after surgery. Likewise, antibody respond may be related to the location of a mature hydatid cyst. Antigen detection shows presence of CE infection, it is extremely important and necessary in follow-up of CE after surgery. The circulating antigen (CAg) titration decrease faster than circulating antibody (during 1–3weeks) after operation. Location of hydatid cyst in detecting antigen is affected less than antibody also. Regarding this subject, antigen detection has several limitations that lead to be used less in CE follow-up. Although, AgB 8/1-kDa subunit is considered as a principle and immunogenic CAg but sensitivity of CAg detection compared to with antibody has variable range, between 33% and 85% which owing to formation of circulating immune complexes (CICs) in result of antigen – antibody complex. The another problem is non using specific CAg (AgB 8/1-kDa subunit) for production of specific paratopes (rabbit hyper immune antiserum) against AgB 8/1-kDa which is used as capture (primary) antibody in double antibody sandwich ELISA assay. The designation of synthetic peptides from conserved regions of AgB 8/1-kDa can help to this problem. These regions (motifs) should be selected for allelic, dominant, immunogenic and conserved without any genetic variation. The first part of this hypothesis suggests which patient’s sera should be treated with acidic buffers such as boric acid, acetic acid, glycine/HCl, polyethylene glycol (PEG) or combination of each of them accompanied by boiling patient’s sera which causes breaking Ag-Ab complexes and in result of releasing AgB 8/1. These modifications are effective to releasing CAg from CIC. To date, the synthetic peptides have been widely used in CE serodiagnosis based on circulating antibody detection only. In second part of this hypothesis suggests the using synthetic peptide of p176 derived AgB 8/1-kDa subunit containing conserved specific epitopes for preparation of specific paratopes (rabbit hyper immune antiserum) based on CAg detection. So, there is no need any native antigens for preparation of non-specific rabbit hyper immune antiserum. These novel improvements can help to decrease the cross-reactivity with other parasitic diseases (specificity). Increasing antigen detection could make a chance in sensitivity of patient’s sera and in result of the best and suitable tool for CE follow-up.

Serological responses in porcine cysticercosis: A link with the parasitological outcome of infection

An oral infection model with Taenia solium whole proglottids was used to study host–parasite relationships and the mechanisms underlying resistance to infection in pigs. In addition, an attempt was made to link the parasitological findings to serological data. Groups of six piglets aged 1, 3 and 5 months were infected and slaughtered 3 months p.i. Circulating antibody and antigen levels were monitored weekly. At autopsy total cyst counts were performed. Although the detailed carcass dissection at necropsy revealed a high variation in the number of cysts, the trend was that the number of viable cysts decreased with the age at which the animals were infected. The kinetics of the antigen levels throughout the course of the infection differed markedly between the three age groups of the experimental infection model. In the younger animals, a fast increase in titres of circulating antigen was observed in most animals, reaching a plateau as early as 2 weeks p.i. Besides its faster increase, antigen levels in pigs infected at younger ages also reached higher levels than in older animals and were associated with weaker antibody responses. Results also demonstrated that a relationship exists between the number of cysts and the titre of circulating antigen. This is promising in view of the development of an assay to quantify the progress of an active T. solium infection and would be a useful tool in epidemiological studies to assess the infection burden and the risk of transmission of the disease. The use of specific antibody-detection assays combined with circulating antigen detection could improve our understanding of this relationship.

Application of a circulating antigen detection immunoassay for laboratory diagnosis of extra-pulmonary and pulmonary tuberculosis

Background Diagnosis of extra-pulmonary tuberculosis is often difficult to establish using standard methods. Recently, a 55-kDa mycobacterial antigen was identified in sera of individuals with pulmonary TB using a simple and rapid dot-ELISA based on monoclonal antibody (TB-55 mAb). Here, we have evaluated the application of the dot-ELISA for the detection of target antigen in sera of individuals with extra-pulmonary TB. Methods The Western blot and indirect immunoperoxidase staining was used to identify the target TB antigen using the TB-55 mAb. The dot-ELISA was used to detect the target antigen in serum samples. Results The target antigen was identified at 55-kDa molecular weight in serum, ascitic fluid and CSF samples from individuals with extra-pulmonary TB. The purified antigen from these samples showed similar biochemical properties to the previously described antigen. The target antigen was localized in areas without caseous necrosis in lymph tissues. The dot-ELISA detected the target antigen in 90% sera of individuals with extra-pulmonary TB and in 87% sera of individuals with pulmonary TB with a specificity of 97% among control individuals. Conclusion The detection of the 55-kDa antigen using dot-ELISA can be routinely employed to support clinical diagnosis of extra-pulmonary TB and pulmonary TB.

Specific and sensitive diagnosis of schistosome infection: can it be done with antibodies?

A simple, cheap, sensitive and specific assay for routine diagnosis of schistosome infection is not yet available. This review considers some of the advantages and disadvantages of parasitological microscopy, circulating-antigen detection and antibody-detection methods. Immunoassays for the detection of antibodies reactive against schistosome egg antigens have the potential to be useful in both clinical and epidemiological settings.

Frequency of canine heartworm disease (D. immitis) in some regions of São Paulo State using a circulating antigen indicator test

The frequency of canine heartworm disease was determined in 310 dogs (158 males and 152 females) from São Paulo city (154) and several towns along the São Paulo state coast (156). The survey was carried out through the Knott's technique (direct presence of microfilariae in the blood) and ELISA for soluble circulating antigen detection. Twenty five (8.0%) samples were positive to D. immitis using both methods being 6 dogs (3.9%) from the city and 19 (12.2%) from the coastal area. Nineteen dogs (6.1%) were positive to the Knott's technique (4 from the city and 15 from the coastal area) and 12 dogs (3.9%) to the ELISA (4 from the city and 8 from the coastal area); six dogs (1.9%) were positive for both methods being two from the city and four from the coastal area. The frequency of positive dogs by the Knott's technique in the coastal area was higher than that in São Paulo city (P<0.05), but equivalent frequency was found in dogs from both areas when the ELISA test was used. Higher number of dogs with a patent infection (microfilaremic) was found than those with occult infection (amicrofilaremic). This survey has shown that the dogs were not infected with high frequency probably because the studied areas were not hiperenzootic endemic (higher than 10% of frequency).

Prévalence de la cysticercose porcine dans le Mayo-Danay (Nord Cameroun) et le Mayo-Kebbi (sud-ouest du Tchad)

Une étude menée de mai à octobre 1999 dans le département du Mayo- Danay (extrême nord du Cameroun) et dans la préfecture du Mayo-Kebbi (sud-ouest du Tchad) a eu pour but de déterminer la prévalence de la cysticercose porcine et d’identifier les principaux facteurs qui la favorisaient. Elle a montré que les conditions hygiéniques dans lesquelles vivait la population ainsi que celles de l’élevage des porcs étaient très médiocres, avec pour conséquence des infestations massives de la population porcine par des cysticerques de Taenia solium. En effet, 42 p. 100 des 126 exploitations visitées étaient dépourvues de latrines et les porcs, en divagation permanente ou semi-permanente, avaient facilement accès aux matières fécales humaines déposées aux alentours des habitations. Le diagnostic clinique effectué par la méthode du langueyage a montré que 20,5 p. 100 des 852 porcs vivants examinés étaient porteurs de cysticerques. L’inspection des carcasses réalisée dans les abattoirs locaux a révélé que 15,7 p. 100 des 51 porcs abattus étaient ladres. Parmi les 264 sérums de porcs soumis au test Elisa pour la détection des antigènes circulants de cysticerques, 105 (39,8 p. 100) se sont révélés positifs. Ces résultats ont indiqué que le Mayo-Danay et le Mayo-Kebbi étaient d’importants foyers de la cysticercose porcine à Taenia solium.

Use of a sandwich-enzyme-linked immunosorbent assay (SEA) for the diagnosis of natural Fasciola hepatica infection in cattle from Galicia (NW Spain)

An indirect-enzyme immune-linked immunosorbent assay (IEA) was compared to a sandwich-enzyme-linked immunosorbent assay (SEA) to diagnose fasciolosis in naturally infected cattle. By means of a rabbit polyclonal IgG antibody to F. hepatica antigens, captured circulating antigens were detected by an SEA technique. A total of 85.1% of the samples examined was considered positive by using the IEA-test, whereas this percentage was of 37.3% by the SEA test. Using the two enzymatic probes, only the 38.3% resulted positive to both immunoassay probes. Our data indicate that circulating antigen detection assay in combination with results of antibody detection may offer a more reliable diagnostic technique than classic coprologic ones, with results very appropriate for epidemiologic studies carried out on a wide area.

Specificity of circulating antigen detection for schistosomiasis mansoni in Senegal and Burundi.

The specificity of schistosome circulating antigen detection was determined in negative individuals from two S. mansoni- endemic countries, Senegal and Burundi, and compared with results from Dutch control individuals. A nearly absolute specificity was achieved for circulating anodic antigen (CAA) detection in serum, irrespective of the target population or sample pretreatment method. Circulating cathodic antigen (CCA) detection in serum and urine resulted in a lower specificity than serum CAA detection. Apparent large differences in specificity of CCA detection between countries were mainly due to pretreatment methods. Apparently, the alkaline/heating pretreatment method is not as effective as trichloroacetic acid (TCA)-pretreatment in removing (certain) interfering components, which may vary between populations. In view of the development of the urine CCA assay into a noninvasive screening test, a slightly lower specificity may still be acceptable. For precise epidemiological analyses the highly specific serum CAA assay remains the method of choice.

Diagnosis of schistosomiasis: antibody detection, with notes on parasitological and antigen detection methods.

Schistosomiasis remains a serious world-wide public health problem with a still unfulfilled need for routine cost-effective methods of diagnosis. Such methods are required not only for people in endemic areas, but increasingly for tourists who may have become infected during visits to such places. This article reviews the wide range of immunoassays and antigenic preparations that have been shown to have potential for diagnosis of schistosomiasis by the indirect method of antibody detection. Antigens in native form derived from cercariae, adult worms and eggs are considered, as well as schistosome antigens produced by recombinant DNA technology and the schistosome cross-reactive antigen, keyhole limpet haemocyanin (KLH). Respective advantages and disadvantages of antibody detection, circulating antigen detection and parasitological methods of diagnosis are analysed. It is suggested that due to the relative insensitivity of both parasitology and antigen detection, antibody detection methods could find increasing use in situations of low infection intensity.

Molecular cloning and characterization of the polypeptide backbone of Schistosomamansoni circulating cathodic antigen

One of the candidate schistosome antigens for the development of a circulating antigen detection diagnostic assay is the circulating cathodic antigen (CCA). Detection of CCA in urine provides a non-invasive assay with high sensitivity. Previously we reported that CCA is secreted in patients’ urine as a small molecular weight material which is probably of a proteinaceous nature. In an attempt to further characterize the secreted component of CCA, we used a monoclonal antibody (MAb) reactive with urine-CCA to isolate an adult worm cDNA clone (SmN3-1) that encodes the polypeptide backbone of CCA. The sequence, gene organization and expression of SmN3-1 were analyzed. The 1.6 kb nucleotide and 347 amino acid sequences of SmN3-1 showed no significant homology to any published sequence. The size and antigenic properties of the expression product of SmN3-1 in Escherichia coli greatly resembled the CCA molecule excreted in urine, suggesting that the latter is primarily composed of the protein element of CCA.

Relevance of circulating antigen detection to follow-up experimental and human cystic hydatid infections.

We analysed specific antibody (Ab) and circulating antigen (CAg) profiles along experimental mouse infection using as control a group of mice immunized with intact but dead parasites. Results from this experiment showed an early major CAg peak followed by a larger Ab peak which partially overlaps with other minor CAg peaks. These results suggest that CAg may be a marker of early mouse infection. In order to study the relevance of these findings in humans we similarly analysed by ELISA 148 sera provided by retrospective post-surgical follow-up of 19 patients. Available records showed that 14 patients developed new cysts one to ten years after surgery while no new disease was observed in the-other five. Some of the former patients showed CAg, as early as two months after surgery while no CAg was observed in the other five patients at any time. In addition, a collection of 38 sera obtained before surgery were similarly tested and five of them showed only CAg, while 18 showed only Ab and 12 sera showed Ab&CAg. These results in humans are consistent with the findings in the mouse experimental model and suggest that CAg may be an early marker of hydatid infection, thus being relevant for post-surgical follow-up.

Diagnosing schistosomiasis.

The ideal diagnostic method for schistosomiasis detection seems to be still far from available. Paucity of egg output in low prevalence situations, low levels of circulating antigens in individuals with low intensity of infection and inadequate specificity of antibody detection systems outline pieces of a puzzle that challenges scientific efforts. Estimated prevalence, financial resources and operational reality must be taken into account when deciding the diagnostic method to be used. A combination of a screening step, using a fast strip test for antibody detection with a parasitological ratification step such as Kato-Katz repeated stool examination may serve as a diagnostic approach for a previously untreated low level endemic area. However, when eradication is the aim, and high financial investment is available, re-treatment may be based on the association between multiple stool examination and circulating antigen detection. Ethical aspects as well as cost-benefit rates between treatment and diagnosis approaches lead to the conclusion that in spite of the recent advances in simple administered and relatively safe drugs, treatment should only be performed when supported by appropriated diagnosis.