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Aims To describe an anti-Strongyloides IgA, IgG and IgG immune complex antibody response profile in patients with pulmonary tuberculosis. Methods and results Saliva and serum samples were collected from 100 individuals: group I, 50 apparently healthy individuals; and group II, 50 pulmonary tuberculosis patients. The IgA, IgG and IgG immune complex detection were carried out via an ELISA immunoenzymatic test. Optical density medians in saliva samples of IgA antibody (median of 7.21) and IgG-IC (median of 4.95) were significantly higher in tuberculosis group compared to control individuals (median IgA of 3.93 and IgG-IC of 2.38). Conclusion This study presents antibody data to the field of pulmonary tuberculosis and strongyloidiasis coinfection, including saliva samples, and especially IgG immune complex detection.

Detection of Strongyloides stercoralis larvae is particularly challenging because only a small number of larvae are released into the feces, regardless of infection stage. Our objective was to apply conventional polymerase chain reaction (PCR) to the detection of S. stercoralis DNA in feces samples to evaluate its performance in samples of patients with strongyloidiasis and compare results with those of immunodiagnosis. Stool, serum, and saliva samples were collected from each individual (n = 48) at the clinic hospital of the State University of Londrina, Brazil, for parasitological, immunological, and molecular tests. Stool samples were processed via parasitological methods. Serum samples were used for immunoglobulin G (IgG) detection and saliva samples for IgA detection by ELISA. For amplification by conventional PCR, two different primers were used: species specific (101 bp) and genus specific (392 bp). The results showed that 34 (97.1%) of the 35 copro-positive individuals for S. stercoralis were positive for serum IgG and 19 (54.3%) were positive for salivary IgA. Regarding molecular analysis, both primers (species and genus specific) demonstrated positivity in 100% of the samples, which was confirmed by sequencing the positive samples. Complementary examinations of the parasitological method demonstrated excellent results in the context of the diagnosis of strongyloidiasis, especially in asymptomatic patients with irregular larval release in the feces.

This review considers the advantages and disadvantages of parasitological techniques, methods of detecting antibodies and antigens, as well as molecular biology techniques in the diagnosis of human strongyloidiasis. In addition, it elucidates the potential of different techniques for rapid and effective detection of clinical cases, thus enabling early treatment and preventing fatal consequences of this helminthiasis.

Human strongyloidiasis is caused by helminthStrongyloides stercoralis. It has a worldwide distribution, often neglected and cause of severe morbidity. The parasitological diagnosis is hindered by the low and irregular amount of larvae in feces. The goal of the present study was to detect IgG and IgG immune complex using conventional serum samples and saliva as alternative samples. We collected samples from 60 individuals, namely: group I composed of 30 healthy individuals; and group II composed of 30 individuals eliminatingS. stercoralislarvae in feces. We calculated the area under the curve, general index of diagnostic accuracy, Kappa index and determined the correlations between different diagnostic tests. The detection of IgG levels was performed by an immunoenzymatic assay with alkaline extract ofS. venezuelensislarvae as antigen. Positivity of anti-S. stercoralisIgG in serum samples from group I was 3·3%, and from group II 93·3%. The detection of immune complex indicated that group I exhibited 3·3% and group II 56·7%. In the saliva samples, IgG detection was 26·7% for group I and 43·3% for group II. Immune complex was detected in 20% of group I, and 30% of group II. IgG immune complex in conventional serum samples and saliva as alternative samples can be considered biomarkers for the diagnosis of active strongyloidiasis.

This study aimed to evaluate the total extract of Taenia crassiceps metacestodes (TC) and its antigenic fractions obtained by Triton X-114 fractionation techniques, such as detergent (DC) and aqueous (AC), in the immunodiagnosis of human neurocysticercosis (NCC). Cerebrospinal fluid samples were divided into two groups: Group 1 (n=40), which was further divided into active (n=20) and inactive (n=20) NCC, and Group 2 (control group), which comprised 39 CSF samples from patients who had another neurological disorder, were suffering from other infectious diseases of the brain or had other parasitic infections. The total extracts and antigenic fractions were tested by enzyme-linked immunosorbent assay (ELISA) to detect human IgG anti-Taenia solium. T. crassiceps fractions (DC and AC) showed the same value of sensitivity (Se), 100%, for active and inactive NCC and a specificity (Sp) of 97.4%. The DS fraction obtained from T. solium showed 100% Se for active NCC, 95% Se for inactive NCC and a 92.3% Sp. The AS fraction obtained from T. solium showed 100% Se for both active and inactive NCC and a 94.9% Sp. There was a positive correlation between the total saline extract of T. crassiceps (TC) and T. solium (TS) and their fractions (DC, AC, DS and AS). Positive predictive value, negative predictive value, diagnostic efficiency and Youden index were calculated. In conclusion, these results demonstrated that detergent and aqueous fractions obtained from T. crassiceps metacestodes are important sources of specific antigens and are efficient for immunodiagnosis of active and inactive NCC.

In experimental infection with Strongyloides venezuelensis, the acute and recovery phases can be distinguished, unlike human infections caused by Strongyloides stercoralis. The objective of this study was to evaluate the production of anti-Strongyloides IgG antibodies and the recognition of immunogenic protein bands during the acute and the recovery phases in rats experimentally infected with S. venezuelensis. Rats were infected subcutaneously with 400 or 4,000 S. venezuelensis infective larvae. The acute phase was characterized by elimination of a large number of eggs in the faeces on days 6-14 post infection; the recovery phase was characterized by the resolution of the infection between days 30 and 35 post infection. Differences in IgG levels were observed in the acute and the recovery phases. Different antigenic fractions were recognized in both phases of infection. It is concluded that proteins within the 30-40 kDa range are immunoreactive markers for both the acute and the recovery phases in rats experimentally infected with S. venezuelensis, particularly using membrane antigen.

Strongyloidiasis is a human parasitic disease caused by the helminth Strongyloides stercoralis whose treatment is particularly difficult in immunosuppressed patients due to their low responsiveness to conventional therapy. Carica papaya and its isolated compounds benzyl isothiocyanate, carpaine and carpasemine are promising compound for the treatment of Strongyloides infections due to their anthelmintic action. This study aims to examine the in vitro ovicidal and larvicidal activity of C. papaya seed hexane extract against Strongyloides venezuelensis, using egg hatching tests and larval motility tests as efficiency markers. The crude extract at the concentrations of 566 – 0.0566 mg/mL or the control with albendazole (0.025 mg/mL) and negative controls (water and PBS) were incubated with an equal volume of egg suspension (± 50 specimens) followed by counting of the specimens after 48 h. The same extract and dilutions were added to L3 larvae suspensions (±50 specimens) followed by analysis of larvae viability after 24, 48, and 72 h. The extract inhibited egg hatching with high efficiency at concentrations of 56.6 mg/mL (95.74%) and 5.66 mg/mL (92.16%). At the concentrations of 566 mg/mL (100%) and 56.66 mg/mL (97.32%), the extract inhibited larval motility as effectively as ivermectin (0.316 mg/mL; 100%), and more effectively than the other dilutions and the negative controls. The larvicidal effect depended on the extract concentration, but not on the treatment period. Therefore, C. papaya seed hexane extract has anthelmintic potential against S. venezuelensis and is a promising compound for the development of phytotherapies to treat strongyloidiasis.

Human ascariasis is a neglected tropical disease of great relevance to public health and is considered the most frequent helminthiasis in poor regions. Accurately diagnosing this parasite has been challenging due to limitations of current diagnostic methods. Immunoglobulin Y (IgY) technology is a very effective alternative for the production of highly specific and profitable antibodies. This study aimed to produce and apply anti-Ascaris suumIgY antibodies in the immunodiagnosis of human ascariasis. Five immunizations comprising total saline extract fromA. suumadult life forms were given at 14-day intervals toGallus gallus domesticushens of the Isa Brown line. Eggs and blood samples were collected weekly and fortnightly, respectively, to monitor the production of antibodies. The specificity of antibodies was confirmed by dot-blot, kinetic enzyme-linked immunosorbent assay (ELISA), avidity ELISA, immunoblotting and indirect immunofluorescence antibody tests. The application for disease diagnosis was performed through the detection of immune complexes in human serum samples by sandwich ELISA. Peaks of IgY anti-A. suumproduction occurred at weeks 6 and 8. IgY showed high avidity levels after the second dose of immunization, ranging from 64% to 93%, with a mean avidity index of 78.30%. Purified IgY recognized 12 bands of proteins fromA. suumsaline extract. Eggs, the uterine portion and cuticles ofA. suumfemale adult are reactive in immunofluorescence. The detection of immune complexes showed diagnostic values of 80% sensitivity and 90% specificity. In conclusion, specific IgY have been shown to be a potential immunodiagnostic tool with promising future applications in human ascariasis.

(1) Background: The validation of biological antigens is the study’s utmost goal in biomedical applications. We evaluated three different probes with single and multiple epitopes through electrochemical detection of specific IgG in serum for human strongyloidiasis diagnosis. (2) Methods: Screen-printed gold electrodes were used and probes consisting of two single-epitope synthetic peptides (D3 and C10) with different sequences, and a multi-epitope antigen [detergent phase (DP)—hydrophobic membrane proteins]. Human serum samples from three populations were used: Strongyloides stercoralis positive, positive for other parasitic infections and negative controls. To test the immobilization of probes onto a screen-printed gold electrode and the serum IgG detection, electrochemical analyses were carried out through differential pulse voltammetry (DPV) and the electrode surface analyses were recorded using atomic force microscopy. (3) Results: The electrochemical response in screen-printed gold electrodes of peptides D3 and C10 when using positive serum was significantly higher than that when using the DP. Our sensor improved sensitivity to detect strongyloidiasis. (4) Conclusions: Probes’ sequences are critical factors for differential electrochemical responses, and the D3 peptide presented the best electrochemical performance for strongyloidiasis detection, and may efficiently substitute whole antigen extracts from parasites for strongyloidiasis diagnosis in electrochemical immunosensors.

SUMMARYThere is an increasing interest in improving neurocysticercosis (NCC) diagnosis through the search of new and alternative antigenic sources, as those obtained from heterologous antigens. The aim of this study was to obtain potential biomarkers for NCC diagnosis after gel filtration chromatography [gel filtration fraction (GFF)] from the total saline extract (SE) from Taenia saginata metacestodes, followed by protein identification and application in immunodiagnostic. SE and GFF proteic profiles were characterized in gel electrophoresis, and diagnostic performance was verified by testing 160 serum samples through enzyme-linked immunosorbent assay and immunoblotting. Sensitivity (Se), specificity (Sp) and other diagnostic parameters were calculated. Polypeptides of interest in the diagnosis of human NCC present at GFF were analysed by mass spectrometry (MS) and B-cell epitopes were predicted. GFF had the best diagnostic parameters: Se 93·3%; Sp 93%; AUC 0·990; LR+ = 13·42 and LR− = 0·07, and proved to be useful reacting with serum samples in immunoblotting. Proteic profile ranged from 64 to 68 kDa and enolase and calcium binding protein calreticulin precursor were identified after MS. The enolase and calcium-binding protein calreticulin precursor showed 18 and 10 predicted B-cell epitopes, respectively. In conclusion we identified important markers in the GFF with high efficiency to diagnose NCC.

Strongyloidiasis is one of the major intestinal infections in humans, and a neglected tropical disease whose diagnosis still poses a challenge. We hypothesized that diagnostic tests based on short peptides containing major epitopes may represent a promising strategy to improve strongyloidiasis detection due to reduced cross-reactivity and higher sensitivity. Our aim was to evaluate two synthetic peptides selected by phage display (C10 and D3) as potential tools for serodiagnosis of strongyloidiasis, and to predict their putative antigen target. To investigate their diagnostic potential, we have tested different panels of serum samples (n = 120) by enzyme linked immunosorbent assay (ELISA) to detect specific IgG, and their diagnostic parameters were calculated. Similarities with proteins from Strongyloides stercoralis were searched and conformational epitopes were predicted and aligned to known protein structures. Both C10 and D3 achieved sensitivity of 95%, and specificities were 89.2% and 92.5%, respectively. D3 presented the highest diagnostic efficiency (93.3%). Epitope prediction for both C10 and D3 led to the alignment with the cytochrome c oxidase subunit 1 structure. In brief, we propose two synthetic peptides as new biomarkers for serodiagnosis of strongyloidiasis, which can be promptly used for ELISA and in future field sensor platforms.

This study evaluates the inclusion of the IgG avidity index in ELISA to detect anti-Strongyloides stercoralis IgG. The ELISA index revealed 70% of specificity. With the inclusion of screening AI, specificity increased to 80%. IgG avidity complemented traditional IgG ELISA by eliminating some of the suspected or false positive cases.

Strongyloides venezuelensis is a model to study human strongyloidiasis, which infects wild rodents and shares common antigenic epitopes with Strongyloides stercoralis. This study aimed to evaluate parasitological and immunological parameters of prednisolone immunosuppression protocols in rats (Rattus novergicus) infected with S. venezuelensis. Rats were divided into six groups (n = 36): untreated and uninfected (-) or infected (+); oral treatment and uninfected (o-) or infected (o+); subcutaneous treatment and uninfected (sc-) or infected (sc+). For oral immunosuppression, 5 mg/mL of water diluted prednisolone were given five days before infection, and in the days 8 and 21 (for 5 days). For subcutaneous immunosuppression, 10 mg/kg of prednisolone were given daily. The infection was established by the subcutaneous injection of approximately 3,000 S. venezuelensis filarioid larvae per animal. All animals from the (+) and (o+) groups survived, while four rats from the (sc+) died prior to necropsy date. Parasitological analysis showed higher egg elimination in (o+) in comparison to (+) and (sc+) on 7, 13 and 26 days post infection (d.p.i.).The recovery of parasitic females at day 30 was significantly higher in (o+), compared to (+). The (+) and (o+) groups showed a clear increase in anti-S. venezuelensis IgG, IgG1 and IgG2 from 13th d.p.i. Oral immunosuppression led to a higher number of adult females and increased egg output while maintaining IgG and subclasses antibody levels comparable to the positive control.

This study aimed to evaluate the use of saliva samples in the Dot-ELISA test for immunodiagnosis of human strongyloidiasis. The Dot-ELISA presented similar results to the ELISA test, with 70% and 60% sensitivity and 85% and 90% specificity, respectively, for IgA in the saliva. The Dot-ELISA with alternative saliva samples may be a suitable tool for diagnosing human strongyloidiasis, especially in populations with high levels of exposure to helminth.

Herein, we evaluate a mimotope-based synthetic peptidenamed NC41 to diagnose neurocysticercosis (NC), a neglected parasitic disease and a major cause of epilepsy worldwide. NC41 synthetic peptide was evaluated to diagnose NC, and total saline extract from Taenia solium metacestodes (SE) was used as control. Serum samples from patients with NC (n = 40), other parasitic diseases (n = 43), and healthy individuals (n = 40) were tested. Diagnostic parameters such as sensitivity (Se), specificity (Sp), likelihood ratio (LR), and area under curve (AUC) were calculated using receiver operating characteristic (ROC) curves. The sequence from T. solium phosphoenolpyruvate carboxykinase (PEPCK) was used for epitope prediction, resulting in one high-scoring patch centered at residue L247. NC41 synthetic peptide reached high diagnostic performance (Se 97.5% and Sp 97.5%, LR+ 39 and AUC 0.997). Data from diagnostic parameters and in silico analyses proved the usefulness of NC41 synthetic peptide as a diagnostic marker for human NC.

Due to the epidemiological problem of the neglected condition of human strongyloidiasis, rapid and effective diagnosis is extremely important, with the development of new diagnostic tools being essential to reduce infections and chronic cases. Avian immunoglobulin Y (IgY) technology is an alternative for antibody production that has high specificity and profitability. This study aimed to produce and fractionate IgY antibodies from the egg yolks of hens that were immunized with the total antigenic extracts of Strongyloides venezuelensis infectious filariform larvae (iL3) and parthenogenetic females (pF). IgY antibodies were then evaluated by their recognition of antigenic proteins, evolutive helminth forms, and serological diagnosis of human strongyloidiasis by the detection of immune complexes in serum samples. Egg yolks were fractionated to obtain IgY antibodies by thiophilic interaction chromatography. Immune complex detection in serum samples showed diagnostic values for anti-iL3 IgY and anti-pF IgY antibodies at 95.56% and 88.89% sensitivity and 95.56% and 91.11% specificity, respectively. Therefore, IgY technology is a promising tool for the detection of blood circulating Strongyloides antigens, with possible application as a serological diagnostic method.

Strongyloidiasis is a human parasitosis that is considered a public health problem. Early diagnosis of this infection is extremely important in immunocompromised patients (i.e. subjects with alcoholism). This study aimed to evaluate anti-Strongyloides immunoglobulin G (IgG) and immunoglobulin A (IgA), assess levels of circulating immune complexes (IC) and determine IgG avidity in serum samples from alcoholic and nonalcoholic individuals. A total of 140 blood samples were collected from male individuals (70 alcoholic and 70 nonalcoholic subjects). Serum was obtained and analysed by enzyme-linked immunosorbent assay for IgG, IgA, IC detection and avidity determination. Anti-Strongyloides IgG was detected in 55.7% of alcoholic subjects and 32.8% nonalcoholics, while IC levels showed frequencies of 38.6% and 17.1% in these groups, respectively. Anti-Strongyloides IgA was lower among alcoholics (4.3%) than nonalcoholics (34.3%). Spearman's correlation coefficient reported a positive correlation between IgG, IC and IgA in alcoholic individuals and no correlation in nonalcoholics. The median avidity index was higher in alcoholics (83.8%) than nonalcoholic subjects (73.2%). In conclusion, this study shows that alcoholic subjects produced specific antibodies against S. stercoralis regardless of the possible immunosuppression caused by chronic alcoholism. Considering that alcoholics are more susceptible to the severe forms of strongyloidiasis, the implementation of immunological methods as a complementary approach to parasitological diagnostics (i.e. detection of IgG, IC and antibody avidity) appears to be an alternative method for early diagnosis in these individuals.

Strongyloidosis is a neglected disease that affects millions of people around the world. The cases that particularly deserve attention are those related to hyperinfection, mainly in immunocompromised patients. In this sense, there is a need to improve the serological diagnosis of this helminth. The objective of this study was therefore to produce and characterize excretory/secretory (E/S) antigens of Strongyloides venezuelensis infective larvae (L3) for use as a heterologous antigen in the diagnosis of human strongyloidosis and other parasitic infection groups. Soluble antigenic preparations were produced as total saline extract (SE), E/S in Roswell Park Memorial Institute 1640 (RPMI) and E/S in phosphate buffered saline (PBS). The three antigenic preparations showed similar protein bands. An ELISA showed that the E/S antigens were profitable, easy to use, and more sensitive and specific than SE, eliminating cross-reactivity with other parasites in serum samples. The detection of anti-Strongyloides stercoralis in the sera of patients with strongyloidosis and those with immunosuppressive conditions using S. venezuelensis L3 larvae E/S antigens was satisfactory. RPMI and PBS E/S antigens were also superior in terms of specificity than SE.

Strongyloides venezuelensis is an important alternative source of antigen for the serologic diagnosis of human strongyloidiasis. Proteomics techniques applied to the analysis of the protein content of infective third stage larvae (iL3) of S. venezuelensis provide a powerful tool for the discovery of new candidates for immunodiagnosis. This study presents an overview of the protein iL3 S. venezuelensis focusing on the diagnosis of strongyloidiasis. A total of 877 proteins were identified by shotgun proteomics. Many of these proteins are involved in different cellular processes, metabolic as well as structural maintenance. Our results point to a catalog of possible diagnostic targets for human strongyloidiasis and highlight the need for evaluation of uncharacterized proteins, especially the proteins within the CAP domain, transthyretin, and BTPI inhibitor domains, as a repertoire as yet unexplored in the context of strongyloidiasis diagnostic markers. We believe that the protein profile presented in this shotgun analysis extends our understanding of the protein composition within the Strongyloides genus, opening up new perspectives for research on biomarkers that may help with the diagnosis of human strongyloidiasis. Data are available via ProteomeXchange with identifier PXD013703.

Human strongyloidiasis is an intestinal helminthiasis that can be fatal particularly in cases of immunosuppression. The aim of this study is to assess the diagnostic accuracy of the detergent fraction (D), purified from total saline extract (SE) of Strongyloides venezuelensis, in the detection of anti-Strongyloides stercoralis IgG antibodies in serum samples from individuals coming from endemic areas for strongyloidiasis and presenting immunocompromised conditions: human immunodeficiency virus (HIV(+)), diabetes mellitus type 2, cancer, tuberculosis and alcoholism. Serum samples from 93 individuals were analyzed by ELISA, as follows: Group 1: 30 immunocompromised individuals with strongyloidiasis; Group 2: 33 immunocompromised individuals without strongyloidiasis and Group 3: 30 healthy individuals. The total saline extract (SE) and detergent fraction (D) showed a sensitivity of 73.33 and 83.33%, and specificity of 82.15 and 86.36%, respectively. The detergent fraction was effective to detect anti-S. stercoralis IgG antibodies in immunocompromised individuals with strongyloidiasis and may be applied as an important tool in the immunodiagnosis of human strongyloidiasis related to immunosuppression.

Human strongyloidiasis is an intestinal parasitosis that may affect 100 million individuals. However, the prevalence rates of this infection may represent smaller values than the actual data, mainly due to difficulties in its diagnosis. The aim of this study was to update the immunological and molecular methods applied to the diagnosis of human strongyloidiasis. There is a great diversity of techniques used in the diagnosis of this parasitosis, such as immunofluorescence antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA), immunoblotting, luciferase immunoprecipitation system (LIPS), dispstick and polymerase chain reaction (PCR), all with advantages and disadvantages, and with unique features for specific purposes. Considering the magnitude of strongyloidiasis and the importance of early diagnosis, due to the possibility of chronicity and hyperinfection, this study analyzes the different methods currently employed, and demonstrates the necessity of developing innovative methodologies, which also maintain diagnostic accuracy, particularly for regions with limited technological resources.

Neurocysticercosis (NC) is one of the most important diseases caused by parasites affecting the central nervous system. We fractionated by ion-exchange chromatography using diethylaminoethyl (DEAE)-sepharose resin the total saline extract (S) from Taenia solium metacestodes and evaluated obtained fractions (DEAE S1 and DEAE S2) by enzyme-linked immunosorbent assay (ELISA, n = 123) and immunoblotting (IB, n = 22) to detect human NC in serum. Diagnostic parameters were established by ROC and TG ROC curves for ELISA tests. IB was qualitatively analyzed. S and DEAE S1 presented sensitivity of 87. 5 % and DEAE S2 90 %. The best specificity was observed for DEAE S2 (90.4 %). In IB, using DEAE S2 samples from NC patients presented bands of 20–25, 43–45, 55–50, 60–66, 82, 89, and 140 kDa. The great diagnostic parameters reached by DEAE S2 suggest the potential applicability of this fraction in NC immunodiagnosis.

We have shown previously that detection of circulating antibodies against mimotopes selected by phage display were useful in neurocysticercosis diagnosis. However, circulating antigens may also be useful in patients’ clinical follow-up. Therefore, we aimed to select novel combinatorial antibodies, single-chain variable fragment (scFv), which can be used for specific antigens with pre-defined affinity and specificity without prior immunization. A phage scFv antibody library was selected against Taenia solium mimotopes displayed on phages coupled in beads and total saline extract of T. solium metacestodes (S) immobilized on microtiter plate wells. After two rounds of selection, 96 phage clones were evolved and validated against each target by enzyme linked immunosorbent assay (ELISA), and dot-blot, and three specific antibodies (B6, G10 and A4) were further characterized by sequencing and indirect immunofluorescence (IFI) assays. IFI revealed tegument staining for the B6, while the others showed a non-uniform staining in the whole parasite. The selected scFvs were used to capture their antigen targets that were elucidated through mass spectrometry, and used for antibody detection in NC patients’ sera by ELISA, which achieved sensitivities greater than 97% and specificities above 95%. We have successfully developed scFv antibodies against important mimotopes used in NC diagnosis, and can be further explored to detect circulating antigens for clinical follow-up of patients with NC. Our strategy also highlighted the possibility of using this combinatorial approach to select, capture and characterize specific antigens to better understand this intriguing parasite infection and disease evolution.

In human and murine models strongyloidiasis induce a Th2 type response. In the current study we investigated the role of different loads of Strongyloides venezuelensis in the immune response raised against the parasite and the participation of the major histocompatibility complex (MHC) class II molecule in the disease outcome in face of the different parasite burden. The C57BL/6 wild type (WT) and MHC II−/− mice were individually inoculated by subcutaneous injection with 500 or 3000 S. venezuelensis L3. The MHC II−/− mice infected with 3000L3 were more susceptible to S. venezuelensis infection when compared with WT groups, in which the parasite was completely eliminated. The production of Th2 cytokines and specific IgG1 or IgE antibodies against parasite were significantly lowered in MHC II−/− infected mice with different larvae inoculums. The infection of MHC II−/− mice with S. venezuelensis induced slight inflammatory alterations in the small intestine, and these lesions were lower when compared with WT mice, irrespective of the parasite load utilized to infect animals. Finally, we concluded that MHC class II molecules are essential in the immune response against S. venezuelensis mainly when infection occurs with high parasite inoculum.

The aim of the present study was to analyse Taenia solium metacestode antigens that were derived from the unbound fraction of jacalin affinity chromatography and subsequent tert-octylphenoxy poly (oxyethylene) ethanol Triton X-114 (TX-114) partitioning in the diagnosis of human neurocysticercosis (NCC). Immunoassays were designed to detect T. solium-specific IgG antibodies by ELISA and immunoblot. Serum samples were collected from 132 individuals who were categorised as follows: 40 had NCC, 62 presented Taenia spp or other parasitic diseases and 30 were healthy individuals. The jacalin-unbound (J unbound ) fraction presented higher sensitivity and specificity rates than the jacalin-bound fraction and only this fraction was subjected to subsequent TX-114 partitioning, resulting in detergent (DJ unbound ) and aqueous (AJ unbound ) fractions. The ELISA sensitivity and specificity were 85% and 84.8% for J unbound , 92.5% and 93.5% for DJ unbound and 82.5% and 82.6% for AJ unbound . By immunoblot, the DJ unbound fraction showed 100% sensitivity and specificity and only serum samples from patients with NCC recognised the 50-70 kDa T. solium-specific components. We conclude that the DJ unbound fraction can serve as a useful tool for the differential immunodiagnosis of NCC by immunoblot.

The aim of this study was to fractionate and partially characterize fractions obtained from the total saline extract (SE) of Taenia saginata metacestodes after ion-exchange procedure in carboxymethyl sepharose (CM) and diethylaminoethyl sepharose (DEAE) resins, as a source of antigenic markers applicable in the immunodiagnosis of neurocysticercosis (NCC). For IgG detection by enzyme-linked immunosorbent assay (ELISA) and immunoblotting, 140 serum samples were analyzed: 45 from patients with NCC (G1), 50 from patients with other parasitic infections (G2), and 45 from healthy individuals. Sensitivity (Se), specificity (Sp), area under curve (AUC), and likelihood ratios (LR) were calculated. CM S2 and DEAE S2 fractions provided high diagnostic values (Se 88.8% and 93.4%; Sp 93.7% and 92.6%; AUC 0.965 and 0.987; LR+ 14.07 and 12.67; LR− 0.11 and 0.07, respectively). In conclusion, CM S2 and DEAE S2 fractions are important sources of specific peptides, with high efficiency to diagnose NCC.

Parasite infections affect billions of people and their domesticated animals worldwide, and remain as a significant cause of morbidity and mortality, but such diseases are still neglected in endemic countries. Therapeutic interventions consisted mostly of drugs, which are highly toxic and may lead to resistance. The immunopathology of parasites is very complex due to their multistage life cycles and long lifetime involving several hosts, leading many times to chronic infections and sometimes to death, by compromising nutritional status, affecting cognitive processes, and inducing severe tissue reactions. Vaccination is a challenge, and immunotherapy is completely disregarded because of their complex interactions with hosts and vectors. This review will bring concepts of immunological aspects for some important parasitic infections, and present the most recent phage display-derived antibodies or peptidomimetics for parasite targets. This chapter will also discuss the future perspectives of such potential anti-infective immunobiologicals for parasitic diseases.

Strongyloides stercoralis is a helminth parasite that can infect millions of people worldwide, particularly in tropical, subtropical and temperate regions with poor sanitation. Several aspects of epidemiology, biology and host-parasite interactions of S. stercoralis have been studied, and substantial knowledge has been acquired; however, very few studies on immunotherapeutic control strategies to prevent infection and disease in humans have been conducted. Therefore, this article reviews the current progress and targets toward vaccine and passive immunization approaches for Strongyloides spp.

This study was performed in order to develop a novel approach based on antigen, antibody and immune complex detection by enzyme-linked immunosorbent assay (ELISA) in bronchoalveolar lavage fluid (BALF) samples. For that purpose Wistar rats immunosuppressed or not were experimentally infected with Strongyloides venezuelensis. The microtiter plates were coated with alkaline parasite extract for antibody detection and with IgG anti-S. venezuelensis for antigen and immune complex detection. The immune serum was able to detect 1.56 μg/mL of L3 antigens in BALF samples. ELISA sensitivity was 96.6%, 71.6% and 91.6% for antigen, antibody and immune complex, respectively, and the specificity was 100% for all methods. Antigen detection in BALF samples showed to be a good approach for evaluating the kinetics of infection in non immunosuppressed or immunosuppressed rats. IgG was detected in non immunosuppressed rats from day 8 p.i. and in immunosuppressed rats from day 2 p.i. Moreover, immune complex was detected during the entire kinetic for both groups. In conclusion, association of antigen, antibody and immune complex detection in BALF samples seems to be an alternative approach for early strongyloidiasis diagnosis particularly in immunosuppressed individuals.

SUMMARYThe aim of this study was to use larval, parasitic female and egg antigens fromStrongyloides venezuelensisto detect parasite-specific IgG and immune complexes in human serum samples by enzyme-linked immunosorbent assay (ELISA). In total, 95 serum samples were analysed, consisting of 30 patients harbouringS. stercoralislarvae, 30 healthy subjects and 35 patients with other parasites. Sensitivity, specificity and diagnostic efficiency were calculated. A significant statistical difference was found in the detection of immune complexes and antibodies in patients harbouringS. stercoralislarvae from larval and eggs antigens, with higher positivity using larval antigen. The larval antigen showed the highest values for sensitivity, specificity and diagnostic efficiency in ELISA from detection of immune complexes. For the first time we used IgG anti-larvae, IgG anti-parasitic females or IgG anti-eggs for immune complex detection. We concluded that the association of antibody and immune complex detection could be used in the diagnosis of human strongyloidiasis.

The aim of this study was to detect levels of IgG and IgA by enzyme-linked immunosorbent assay (ELISA) using alkaline extracts of larvae, adult female worms, and eggs of Strongyloides venezuelensis as antigen. One hundred twenty serum samples divided into 3 groups were analysed: group I (40 strongyloidiasis patients), group II (40 patients with other parasitic infections), and group III (40 healthy subjects). Statistical variations were analyzed using analysis of variance. There was a significant statistical difference (P < 0.001) in the detection of antibodies in group I between larvae and female antigens and between larvae and egg antigens, with higher positivity using larvae antigen. The larvae antigen showed the highest values for sensitivity, specificity, and diagnostic efficiency in ELISA. This study is the first that examines the use of adult female worm and egg antigens to detect antibodies for human strongyloidiasis diagnosis compared with the larval extract. By comparing all 3 extracts, larval antigens demonstrated better diagnostic parameters.

SUMMARYThe objective of this review was to outline an epidemiological profile ofStrongyloides stercoralisby parasitological and serological diagnosis in inhabitants, and to associate this profile with different immunosupression situations, in Brazil, over 20 years (1990–2009). The occurrence ofS. stercoralisusing parasitological methods was 5·5%, being 4·8% in rural and 5·0% in urban areas, characterizing the country as hyperendemic. There was a diversity of techniques used as a diagnostic tool and only 39·1% of the studies presented results based on at least 1 specific method. The occurrence increased with age, being 12·1%, for those over 60 that suggests an epidemiological condition of concern for the elderly population. Of the seroepidemiological studies in the general population the mean positivity in serum samples was 21·7% and 29·2%, using an immunofluorescence antibody test and enzyme-linked immunosorbent assay (ELISA), respectively. The occurrence of strongyloidiasis in immunosuppressed individuals was 11·8% by parasitological methods and 19·5% using immunological methods. Considering that Brazil is a tropical country and that the character of chronicity and autoinfection of the parasite that can result in severe forms of hyperinfection or dissemination makes strongyloidiasis an important medically and socially neglected problem.

The aim of this study was to investigate the immunomodulatory effects of glucocorticoids on the immune response to Strongyloides venezuelensis in mice. Balb/c mice were infected with S. venezuelensis and treated with Dexamethasone (Dexa) or vehicle. Dexa treatment increased circulating blood neutrophil numbers and inhibited eosinophil and mononuclear cell accumulation in the blood, bronchoalveolar, and peritoneal fluid compared with control animals. Moreover, Dexa decreased tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-3 (IL-3), IL-4, IL-5, IL-10, and IL-12 production in the lungs and circulating immunoglobulin G1 (IgG1), IgG2a, and IgE antibody levels while increasing the overall parasite burden in the feces and intestine. Dexa treatment enhanced the fertility of female nematodes relative to untreated and infected mice. In summary, the alterations in the immune response induced by Dexa resulted in a blunted, aberrant immune response associated with increased parasite burden. This phenomenon is similar to that observed in S. stercoralis-infected humans who are taking immunosuppressive or antiinflammatory drugs, including corticosteroids.

Summary Neurocysticercosis (NC), caused by Taenia solium metacestode, infects the central nervous system and is a devastating parasitic infection. Diagnosis is based on symptoms, imaging, serology and epidemiology. Current markers present variable sensitivity and specificity, frequent cross-reactions and are not able to discriminate NC clinical forms. The aim of this study was to select mimotopes of T. solium metacestode antigens that may be used in NC immunodiagnosis, specifically to discriminate between active and inactive forms. A random peptide phage display library was screened against IgY from chickens immunized with total saline extract from T. solium metacestodes and validated against 110 serum samples, classified into active NC (18), inactive NC (22), cross-reactive parasitic diseases (40) and healthy controls (30). We have successfully selected seven peptides with significant immunoreactivity to IgG of NC patients, with sensitivity ranging from 95·5% to 100% to detect the inactive form and specificity varied from 85·7% to 94·3%. One phage-displayed peptide (Cc48) can be directly used as biomarker to distinguish inactive from active forms with an accuracy of 95·7%, and this novel mimotope may also be used as an auxiliary tool to neuroimaging tests and treatment follow-up.

The present research investigated the influence of temperature and time of larvae culture on the infectivity of Strongyloides venezuelensis. Mice were infected s.c. with 1500 larvae of S. venezuelensis maintained at 28 °C for three days of culture (dc), 28 °C for seven dc or 18 °C for seven dc. On days 1, 3, 5, 7, 14 and 21 post-infection the animals were sacrificed and cell numbers in the blood, peritoneal cavity fluid (PCF), broncoalveolar fluid (BALF), cytokines, immunoglobulins, number of parasites and eggs/g of feces were quantified. Results demonstrated an increase in eosinophils and mononuclear cells in the blood, PCF and BALF of infected mice. Larvae at 28 °C/3dc induced earlier eosinophils in the PCF and BALF as opposed to larvae at 28 °C/7dc and 18 °C/7dc. Larvae at 28 °C/7dc induced higher synthesis of IL-4, IL-5 and IL-10 on days 5 and 7 post-infection. Larvae at 28 °C/3dc in culture induced higher synthesis of IL-12 than larvae of seven dc, but time in culture induced better synthesis of IFN-γ after larval migration had ceased and only adult worms were present. Larvae at 28 °C/3dc in culture induced higher synthesis of IgG and IgG1 and expelled less female parasites than larvae cultivated for seven days. In conclusion, it was observed that the infectivity of S. venezuelensis is influenced by variations in temperature and time of culture.

The aim of this study was to evaluate jacalin-bound fraction (JBF) and jacalin-unbound fraction (JUF) of the total saline extract from Taenia saginata metacestodes for human neurocysticercosis (NC) immunodiagnosis in cerebrospinal fluid. Total extract, JBF, and JUF were separated by affinity chromatography using Sepharose(®)-jacalin and were tested in enzyme-linked immunosorbent assay (ELISA) and Western blotting (WB) to detect immunoglobulin G. In ELISA test, JUF showed the higher diagnostic efficiency and specificity indexes, 92% and 100%, respectively. In WB, 5 immunodominant proteins (39-42, 47-52, 64-68, 70, and 75 kDa) were detected when using JUF. In conclusion, the results achieved demonstrate that JUF, obtained from T. saginata metacestodes, are an important source of specific peptides and are efficient in the diagnosis of NC.

The objective of the present research was to evaluate detergent and aqueous phases of total saline (TS) and alkaline extracts of Strongyloides venezuelensis for human strongyloidiasis immunodiagnosis. Total extracts and detergent and aqueous antigenic fractions were separated using Triton X-114 and were examined by enzyme-linked immunosorbent assay (ELISA) and immunoblotting (IB) tests to detect immunoglobulin G (IgG). Serum samples were obtained from 120 individuals: 40 strongyloidiasis patients (group I), 40 patients with other parasitic diseases (group II), and 40 apparently healthy individuals (group III). Each extract provided a different profile of antigenic components as recognized by IgG in IB. The detergent fraction of the TS extract demonstrated the highest sensitivity and specificity for ELISA and IB. The results indicated that the detergent saline fraction, purified from S. venezuelensis, furnished the most valid results for the strongyloidiasis immunodiagnosis and could be employed as an alternative antigen and as a useful source of specific polypeptides.

Taenia saginata metacestode antigens have been constituted a useful alternative antigen for neurocysticercosis (NC) serodiagnosis, particularly due to an increasing difficulty to obtain Taenia solium homologous antigen. Cross-reactivity with Echinococcus granulosus infection occurs in homologous and heterologous antigens and could be avoided by using different purified methods. The present study evaluated antigen fractions obtained from saline extracts of T. saginata metacestodes purified by affinity chromatography with jacalin or concanavalin A (ConA) lectins to detect IgG antibodies by enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis to diagnose human NC. Serum samples were collected from 142 individuals: 40 of them were diagnosed with NC, 62 presented Taenia sp. and other parasites, and 40 were apparently healthy individuals. The jacalin- and ConA-unbound fractions demonstrated sensitivity and specificity higher than those of bound fractions. Among unbound fractions, ConA demonstrated statistically higher sensitivity and specificity by ELISA (90% and 93.1%, respectively). By immunoblot assay, the 64- to 68-kDa component from the ConA-unbound fraction showed 100% sensitivity and specificity, making this component suitable for use as a specific antigen for diagnosis of NC. To our knowledge, this is the first report showing the relevance of using the unbound ConA fraction of T. saginata metacestodes to diagnose NC. In conclusion, the results obtained herein clearly demonstrate that antigenic fractions without affinity to ConA, obtained from T. saginata metacestodes, are an important source of specific peptides and are efficient in the diagnosis of NC when tested by immunoblot assay.

Neurocysticercosis (NC), caused by Taenia solium, is the most common infection caused by helminthes of the human central nervous system. In this study, a random peptide phage display library was used to isolate peptide ligands as potential markers for neurocysticercosis diagnosis, because occurrence of cross-reactions with other helminthes species in the current used markers. We selected different peptides using IgG purified from pooled sera of neurocysticercosis patients. To investigate the diagnostic potential of recombinant peptides, we have tested different panels of serum samples by Phage-ELISA, and 10 phage clones strongly bound to the anti-T. solium IgGs in NC sera, with an accuracy range from 84.2% to 95%. The phage clones, NC(4)1 and NC(2)8, presented the highest sensitivity and specificity (100%), respectively, and most important, some phage clones did not react with patients' sera from Echinococcus granulosus infected patients. The validation with a competitive ELISA assay demonstrated that the selected phages could mimic T. solium epitopes and bind specifically to the pool of NC sera. Finally, the two recombinant antigens may become potential biomarkers for serodiagnosis of NC, and the Phage-ELISA demonstrated to be a very good assay, being reproducible, simple, fast, and low-cost due to its production through Escherichia coli culture, allowing a high throughput screening of NC.

INTRODUCTION: from an epidemiological point of view, more than 120 species of triatomine (Hemiptera, Reduviidae) are known. The occurrence and positivity for Trypanosoma cruzi in triatomines in 16 municipalities of the Triângulo Mineiro and Alto Paranaíba were evaluated from January 2002 to December 2004. METHODS: the triatomines were captured basically according to the classic norms of the National Health Foundation. The parasitological exams of the triatomines were conducted according to the technique described by the Ministry of Health. During the study period, 990 specimens of triatomines were captured and of these, 771 could be examined. RESULTS: five species were identified: Triatoma sordida, Panstrongylus diasi, Panstrongylus megistus, Panstrongylus geniculatus and Rhodnius neglectus. Triatoma sordida represented 71.5% of all the triatomines captured, followed by Panstrongylus megistus (18%), Rhodnius neglectus (9.3%), Panstrongylus diasi (0.8%) and Panstrongylus geniculatus (0.4%). Of the total number of triatomines examined, 2.7% were positive for Trypanosoma cruzi. Panstrongylus megistus was the species that presented the highest rates of infection by Trypanosoma cruzi (8.3%), followed by Rhodnius neglectus (2.9%) and Triatoma sordida (1.4%). CONCLUSIONS: there is a need to adapt to new circumstances in epidemiology, with greater emphasis on entomological surveillance, since the potential for adaptation of secondary species of triatomines exists, especially where Chagas' disease is already under control. INTRODUÇÃO: do ponto de vista epidemiológico mais de 120 espécies de triatomíneos (Hemiptera, Reduviidae) são conhecidas. A ocorrência e a positividade de Trypanosoma cruzi em triatomíneos de 16 municípios do Triângulo Mineiro e Alto Paranaíba foram avaliadas de janeiro de 2002 a dezembro de 2004. MÉTODOS: os triatomíneos foram capturados seguindo basicamente as normas clássicas da Fundação Nacional de Saúde. Os exames parasitológicos dos triatomíneos foram conduzidos de acordo com a técnica descrita pelo Ministério da Saúde. Durante o período de estudo, foram capturados 990 exemplares de triatomíneos, sendo que 771 dos capturados estavam em condições de serem examinados. RESULTADOS: cinco espécies foram identificadas: Triatoma sordida, Panstrongylus diasi, Panstrongylus megistus, Panstrongylus geniculatus e Rhodnius neglectus. Triatoma sordida representou 71,5% de todos os triatomíneos capturados, seguido por Panstrongylus megistus (18%), Rhodnius neglectus (9,3%), Panstrongylus diasi (0,8%) e Panstrongylus geniculatus (0,4%). Dos triatomíneos examinados, 2,7% foram positivos para Trypanosoma cruzi. Panstrongylus megistus foi a espécie que apresentou a maior taxa de infecção por Trypanosoma cruzi (8,3%), seguida pelo Rhodnius neglectus (2,9%) e Triatoma sordida (1,4%). CONCLUSÕES: há necessidade de se adequar às novas circunstâncias epidemiológicas com ênfase na vigilância entomológica, uma vez que o potencial de adaptação de espécies secundárias de triatomíneos, em áreas onde a doença de Chagas está controlada, é uma preocupação.

Strongyloides stercoralis is an intestinal nematode capable of chronic, persistent infection and hyperinfection of the host; this can lead to dissemination, mainly in immunosuppressive states, in which the infection can become severe and result in the death of the host. In this study, we investigated the immune response against Strongyloides venezuelensis infection in major histocompatibility complex (MHC) class I or class II deficient mice. We found that MHC II(-/-) animals were more susceptible to S. venezuelensis infection as a result of the presence of an elevated number of eggs in the faeces and a delay in the elimination of adult worms compared with wild-type (WT) and MHC I(-/-) mice. Histopathological analysis revealed that MHC II(-/-) mice had a mild inflammatory infiltration in the small intestine with a reduction in tissue eosinophilia. These mice also presented a significantly lower frequency of eosinophils and mononuclear cells in the blood, together with reduced T helper type 2 (Th2) cytokines in small intestine homogenates and sera compared with WT and MHC I(-/-) animals. Additionally, levels of parasite-specific immunoglobulin M (IgM), IgA, IgE, total IgG and IgG1 were also significantly reduced in the sera of MHC II(-/-) infected mice, while a non-significant increase in the level of IgG2a was found in comparison to WT or MHC I(-/-) infected mice. Together, these data demonstrate that expression of MHC class II but not class I molecules is required to induce a predominantly Th2 response and to achieve efficient control of S. venezuelensis infection in mice.

Strongyloidiasis, caused by the nematode Strongyloides stercoralis, is one of the major worldwide parasitic infections in humans. Breastfeeding may offer a potential protection against this infection. Feces, serum and milk samples were obtained from 90 lactating women from Clinical Hospital of Universidade Federal de Uberlândia, Brazil. The fecal samples were collected for parasitological diagnosis and the serum and milk samples were examined for specific S. stercoralis IgA and IgG antibodies using the indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). Fecal examination showed that the rate of prevalence of S. stercoralis infection in the lactating women was 4.4%. IFAT manifested a 16.7% positivity rate for specific IgA antibody in serum and a 28.9% rate in milk samples; specific IgG was 41.1% in serum and 25.5% in milk samples. According to ELISA the positivity rate for specific IgA antibody was 21.1% in serum and 42.2% in milk samples; specific IgG was 40% in serum and 18.9% in milk samples. In serum samples, these immunological tests showed a concurrence of 91.1% and 94.4%, respectively, in detecting specific IgA and IgG antibodies. In milk samples, they showed a concurrence of 70% and 78.9%, respectively, in detecting specific IgA and IgG antibodies. There was a statistically significant difference between concordant and discordant results of immunological tests (P

The aim of this study was to determine the occurrence of intestinal parasites and commensals among children in four peripheral districts located in the northern, southern, eastern and western sectors of Uberlândia, Minas Gerais, using the Baermann methods as modified by Moraes and Lutz. Out of 160 individuals studied, 93 (58.1% CI: 50.4-65.7) were infected, distributed among the sectors as follows: northern (72.5%), southern (47.5%), eastern (57.5%) and western (55%). The positive findings according to age groups were: 0-5 years (26.9%), 5-10 years (21.2%) and 10-15 years (10%). Male children presented 2.7 times higher risk of infection than females did (OR: 2.7; CI: 1052-7001). The parasites and commensals identified were: Giardia lamblia (27.5%), Entamoeba coli (20.6%), Ascaris lumbricoides (14.4%), Enterobius vermicularis (8.8%), Hymenolepis nana (7.5%), Hymenolepis diminuta (5%), hookworms (3.1%), Trichuris trichiura (2.5%), Endolimax nana (2.5%), Entamoeba hartmanni (2.5%), Strongyloides stercoralis (1.3%), Iodamoeba butschlii (1.3%) and Capillaria hepatica (0.6%). The infection rate in these children was high and showed the need to implement prophylactic education programs in the community.

The present study, investigated the mechanisms involved in the immune responses of Major Histocompatibility Complex class I or class II knockout mice, following Strongyloides venezuelensis infection. Wild-type C57BL/6 (WT), MHC II −/− and MHC I −/− mice were individually inoculated with 3000 larvae (L3) of S. venezuelensis and sacrificed on days 1, 3, 5, 8, 13 and 21 post-infection (p.i.). Samples of blood, lungs and small intestines were collected. The tissue samples were stained with hematoxylin–eosin for the pathological analysis. The presence of the parasite was demonstrated by immunoperoxidase analysis. MHC II −/− mice presented a significantly higher number of adult worms recovered from the small intestine on day 5 p.i. and presented elevated numbers of eggs in the feces. The infection by S. venezuelensis was completely eliminated 13 days after infection in WT as well as in MHC I −/− mice. In MHC II −/− mice, eggs and adult worms were still found on day 21 p.i., however, there was a significant reduction in their numbers. In the lung, the parasite was observed in MHC I −/− on day 1 p.i. and in MHC II −/− mice on days 1 and 5 p.i. In the small intestine of WT mice, a larger number of parasites were observed on day 8 p.i. and their absence was observed after day 13 p.i. Through immunohistochemistry analysis, the parasite was detected in the duodenum of WT on days 5 and 8 p.i., and in knockout mice on days 5, 8 and 13 p.i.; as well as in posterior portions of the small intestine in MHC I −/− and MHC II −/− on day 13 p.i., a finding which was not observed in WT mice. We concluded that immunohistochemistry analysis contributed to a more adequate understanding of the parasite localization in immunodeficient hosts and that the findings aid in the interpretation of immunopathogenesis in Strongyloides infection.

OBJECTIVE To evaluate the detergent and aqueous phases of crude saline extract (crude) of Taenia solium metacestodes extracted with Triton X-114, for the detection of immunoglobulin G (IgG) antibodies by enzyme-linked immunosorbent assay (ELISA) and immunoblot for the laboratory diagnosis of human neurocysticercosis. METHOD Serum samples were collected from 138 individuals and tested; 40 were diagnosed with neurocysticercosis (NC; group 1), 58 presented other parasitic diseases (group 2) and 40 were apparently healthy (group 3). RESULTS The detergent phase showed only one dominant component (70-50 kDa), while the aqueous phase showed four (116, 110, 97 and 77 kDa) by silver staining. ELISA sensitivity and specificity were 92.5% and 84.5% for crude phase, 92.5% and 93.3% for detergent phase and 72.5% and 72.6% for aqueous phase. CONCLUSIONS The immunoblot of the antigens confirmed the results obtained by ELISA. Detergent phase purified from T. solium metacestodes was an important source of specific peptides and was efficient in detecting T. solium metacestodes antibodies in serum samples. This phase can be used for detecting IgG antibodies to T. solium metacestodes in seroepidemiological investigations and in the diagnostic screening of NC patients.

The aim of this study was to evaluate total IgG, IgG1, IgG4, and IgE antibody responses in human strongyloidiasis by enzyme-linked immunosorbent assay (ELISA) using Strongyloides ratti saline extract as heterologous antigen for a possible clinical utility of the assay. A total of 40 serum samples of patients who were shedding Strongyloides stercoralis larvae in feces (group I), 30 sera from patients with other intestinal parasites (group II), and 30 sera from subjects with negative results in three parasitological assays (group III) were analyzed to detect total IgG, IgG1, IgG4, and IgE to Strongyloides spp. by ELISA and expressed in ELISA index. Levels of total IgG anti-Strongyloides spp. were significantly higher in patients of group I than in groups II (p = 0.0005) and III (p < 0.0001). Levels of specific IgG1, IgG4, and IgE of group I were also significantly higher than in groups II and III, respectively. There was a significant positive correlation between specific IgE and IgG4 (r = 0.6524; p = 0.0084) and IgG1 and IgG4 (r = 0.5398; p = 0.0171). It can be concluded that the detection of specific IgE, IgG1, and IgG4 subclasses rather than total IgG antibodies to Strongyloides spp. using the S. ratti antigen showed to be an additional tool for improving the serodiagnosis of human strongyloidiasis.

SUMMARYDefinitive diagnosis of strongyloidiasis in humans is typically achieved by detection of larvae in fecal samples. However, limitations on sensitivity of parasitological methods emphasize the need for more robust diagnostic methods. The aim of this study was to compare the diagnostic value of three methods: eggs per gram of feces (EPG), coproantigen detection by enzyme linked immunosorbent assay (ELISA), and DNA detection by conventional polymerase chain reaction (PCR). The assays were performed at 0 and 5, 8, 13, 21 and 39 days post-infection (dpi) using fecal samples from experimentally infected immunocompetent and immunosuppressed rats. In immunocompetent rats, eggs were detected in feces on days 5, 8 and 13 dpi; coproantigen detection and PCR amplification were successful at all post-infection time points (5, 8, 13, 21 and 39 dpi). In immunosuppressed rats, eggs were detected at 5, 8, 13 and 21; coproantigen detection and PCR amplification were successful at all post-infection time points. In conclusion, these results suggest that coproantigen detection and PCR may be more sensitive alternatives to traditional methods such as EPG for diagnosis of Strongyloides venezuelensis infection.