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1.
J Helminthol ; 94: e71, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31409433

RESUMO

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 suum IgY antibodies in the immunodiagnosis of human ascariasis. Five immunizations comprising total saline extract from A. suum adult life forms were given at 14-day intervals to Gallus gallus domesticus hens 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. suum production 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 from A. suum saline extract. Eggs, the uterine portion and cuticles of A. suum female 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.


Assuntos
Anticorpos Anti-Helmínticos/biossíntese , Ascaríase/diagnóstico , Ensaio de Imunoadsorção Enzimática , Imunoglobulinas/biossíntese , Animais , Complexo Antígeno-Anticorpo , Antígenos de Helmintos/administração & dosagem , Antígenos de Helmintos/imunologia , Ascaris suum , Galinhas , Feminino , Humanos , Imunização , Testes Imunológicos/métodos , Sensibilidade e Especificidade
2.
Parasitology ; 145(8): 1090-1094, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29239289

RESUMO

Human strongyloidiasis is caused by helminth Strongyloides 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 eliminating S. stercoralis larvae 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 of S. venezuelensis larvae as antigen. Positivity of anti-S. stercoralis IgG 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.


Assuntos
Complexo Antígeno-Anticorpo/análise , Antígenos de Helmintos/imunologia , Imunoglobulina G/análise , Testes Imunológicos/métodos , Saliva/química , Estrongiloidíase/diagnóstico , Animais , Anticorpos Anti-Helmínticos/sangue , Complexo Antígeno-Anticorpo/sangue , Biomarcadores/análise , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Humanos , Imunoglobulina G/sangue , Larva , Strongyloides stercoralis/imunologia , Estrongiloidíase/imunologia
3.
Parasitology ; 144(4): 426-435, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-27748211

RESUMO

There 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.


Assuntos
Cromatografia em Gel/métodos , Proteínas de Helminto/metabolismo , Neurocisticercose/sangue , Neurocisticercose/diagnóstico , Taenia saginata/metabolismo , Animais , Biomarcadores/sangue , Fracionamento Químico , Ensaio de Imunoadsorção Enzimática , Epitopos de Linfócito B , Proteínas de Helminto/sangue , Proteínas de Helminto/genética , Humanos , Programas de Rastreamento , Modelos Moleculares , Neurocisticercose/parasitologia , Conformação Proteica , Taenia saginata/isolamento & purificação
4.
J Helminthol ; 90(1): 117-20, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25327496

RESUMO

This study aimed to compare three qualitative parasitological methods for the diagnosis of Syphacia muris infection in 30 Wistar rats (Rattus norvegicus) infected naturally. Methods of spontaneous sedimentation (Hoffman, Pons and Janer, or HPJ) and spontaneous flotation (Willis) for faecal samples and a method of taping (Graham) were performed and compared. The Graham and Willis methods were more sensitive than the HPJ method (P< 0.05). The Graham method was able to detect S. muris eggs in 100% of the samples. Eggs were detected in 83% and 60% of the samples using the Willis and HPJ methods, respectively. Method choice is important for screening for parasites of rats kept under laboratory conditions, as accurate diagnosis helps prevent future environmental contamination and infection. We concluded that the Graham method was the most efficient of those tested in this study for detection of S. muris infection in rats. This method is also rapid, inexpensive and practical, and should be implemented as a necessary measure for infection control.


Assuntos
Oxiuríase/veterinária , Oxyuroidea/isolamento & purificação , Parasitologia/métodos , Doenças dos Roedores/diagnóstico , Animais , Feminino , Masculino , Oxiuríase/diagnóstico , Oxiuríase/parasitologia , Oxyuroidea/fisiologia , Ratos , Ratos Wistar , Doenças dos Roedores/parasitologia
5.
J Helminthol ; 90(4): 422-7, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26169305

RESUMO

Strongyloides venezuelensis is an intestinal nematode of rats, frequently used as a model for studying human and animal strongyloidiasis. In the present study, we evaluated parasitological, serological and molecular methods for the diagnosis of experimental S. venezuelensis in rats, Rattus norvegicus. Blood and faecal samples were collected and analysed up to 60 days post infection (pi) with adult worm recovery occurring from 5 to 45 days pi. Using an enzyme-linked immunosorbent assay (ELISA), serum levels of IgG antibodies increased up to 28 days pi, thereafter decreasing by day 60 pi. Polymerase chain reaction (PCR) assays detected S. venezuelensis DNA in faecal samples of rats from 5 to 21 days pi. The present study therefore represents the first step towards improving the diagnosis of experimental strongyloidiasis.


Assuntos
Testes Diagnósticos de Rotina/métodos , Strongyloides/isolamento & purificação , Estrongiloidíase/diagnóstico , Animais , Sangue/parasitologia , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Técnicas de Diagnóstico Molecular/métodos , Parasitologia/métodos , Reação em Cadeia da Polimerase , Ratos , Testes Sorológicos/métodos
6.
J Helminthol ; 90(3): 342-6, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-25997825

RESUMO

In the present study, antigens from parthenogenetic females and eggs of Strongyloides venezuelensis, or anti-parthenogenetic-female and anti-egg antigens were used to detect specific IgG and immune complex responses, respectively. Serum samples from experimentally infected immunocompetent and immunosuppressed rats were analysed on days 5, 8, 13 and 21 post-infection (dpi). An enzyme-linked immunosorbent assay (ELISA) was performed using alkaline parasite extract for specific IgG detection, and anti-parthenogenetic-female or anti-egg antigens for immune complex detection. The data were analysed using analysis of variance (ANOVA), followed by a Bonferroni test. When parthenogenetic female or egg extracts were used as antigens, specific IgGs were not detected in either immunocompetent or immunosuppressed rats. When anti-parthenogenetic-female or anti-S. venezuelensis-eggs were used, immune complexes were detected for the duration of the infection in immunosuppressed animals and were only detected between 5 and 13 dpi in immunocompetent animals. The duration of infection was not significantly different between the immunocompetent and immunosuppressed groups when anti-parthenogenetic-female or anti-S. venezuelensis-eggs were used. Parthenogenetic female extracts yielded significant differences between antibody and immune complex responses in immunocompetent rats from 5 to 13 dpi, but only on day 5 dpi in immunosuppressed rats. Exposure to S. venezuelensis egg extract yielded significant differences in both antibody and immune complex detection between immunocompetent and immunosuppressed rats for the duration of the infection. In conclusion, ELISA using alternative antigens may be a successful strategy for identifying immune complexes in serum samples and diagnosing active strongyloidiasis, particularly under conditions of immunosuppression.


Assuntos
Anticorpos Anti-Helmínticos/sangue , Complexo Antígeno-Anticorpo/sangue , Imunoglobulina G/sangue , Terapia de Imunossupressão , Strongyloides/imunologia , Estrongiloidíase/diagnóstico , Zigoto/imunologia , Animais , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Masculino , Ratos , Estrongiloidíase/imunologia , Estrongiloidíase/parasitologia
7.
Parasitology ; 140(1): 69-75, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22931968

RESUMO

The aim of this study was to fractionate and partially characterize the antigenic extract of filariform larvae of Strongyloides venezuelensis in ion-exchange resin diethylaminoethyl sepharose (DEAE), to obtain antigenic fractions potentially applicable in immunoassays. Somatic antigen (SA) and its fractions DEAE S1 and DEAE S2 - which interacted with the resin - were evaluated by 1-dimensional electrophoresis to obtain protein profiles. SA and its fractions were tested in serum samples for IgG detection by ELISA. Serum samples (n = 155) were analysed: 50 from strongyloidiasis patients (G1), 55 from patients with other parasitic infections (G2) and 50 from healthy volunteers. Sensitivity (Se), specificity (Sp), area under curve (AUC) and likelihood ratios (LR) were calculated. The DEAE S2 fraction provided a high diagnostic value for IgG detection (Se 92·0%, Sp 91·4%, AUC 0·981, LR+ 10·75, LR - 0·09). In conclusion, the DEAE S2 fraction would probably be a source of immunodominant polypeptides for IgG detection in human strongyloidiasis serodiagnosis.


Assuntos
Antígenos de Helmintos , Cromatografia por Troca Iônica , Strongyloides/química , Estrongiloidíase/diagnóstico , Animais , Ensaio de Imunoadsorção Enzimática/normas , Humanos , Imunoglobulina G/sangue , Larva/química , Sensibilidade e Especificidade , Testes Sorológicos , Soro/parasitologia
8.
J Helminthol ; 87(3): 301-4, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22776357

RESUMO

Little is known about the actin cytoskeleton architecture in female Strongyloides venezuelensis and thus to investigate the distribution and concentration of actin, female worms were labelled with phalloidin-rhodamine and visualized under confocal microscopy. Our results demonstrate that filamentous actin accumulates in the vulva and the concentration of F-actin at this site suggests its important role, especially during oviposition, in the life cycle of S. venezuelensis.


Assuntos
Actinas/análise , Strongyloides/química , Animais , Feminino , Microscopia Confocal/métodos , Oviposição , Coloração e Rotulagem/métodos , Strongyloides/fisiologia , Vulva/química
9.
Parasitology ; 139(7): 956-61, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22309625

RESUMO

The aim of this study was to use larval, parasitic female and egg antigens from Strongyloides venezuelensis to 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 harbouring S. stercoralis larvae, 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 harbouring S. stercoralis larvae 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.


Assuntos
Complexo Antígeno-Anticorpo/sangue , Antígenos de Helmintos , Imunoglobulina G/sangue , Strongyloides/imunologia , Estrongiloidíase/diagnóstico , Animais , Especificidade de Anticorpos , Antígenos de Helmintos/imunologia , Feminino , Humanos , Testes Imunológicos , Larva/imunologia , Óvulo/imunologia , Ratos , Ratos Wistar , Sensibilidade e Especificidade , Strongyloides/classificação , Estrongiloidíase/imunologia , Estrongiloidíase/parasitologia
10.
Parasitology ; 138(11): 1331-40, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21810305

RESUMO

The objective of this review was to outline an epidemiological profile of Strongyloides stercoralis by parasitological and serological diagnosis in inhabitants, and to associate this profile with different immunosupression situations, in Brazil, over 20 years (1990-2009). The occurrence of S. stercoralis using 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.


Assuntos
Anticorpos Anti-Helmínticos/sangue , Strongyloides stercoralis/fisiologia , Estrongiloidíase , Adolescente , Adulto , Fatores Etários , Idoso , Animais , Anticorpos Anti-Helmínticos/imunologia , Brasil/epidemiologia , Criança , Pré-Escolar , Bases de Dados Bibliográficas , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Hospedeiro Imunocomprometido , Imunoglobulina G/sangue , Masculino , Pessoa de Meia-Idade , Prevalência , Sensibilidade e Especificidade , Estudos Soroepidemiológicos , Testes Sorológicos , Estrongiloidíase/diagnóstico , Estrongiloidíase/epidemiologia , Estrongiloidíase/imunologia , Estrongiloidíase/parasitologia
11.
Parasite Immunol ; 33(6): 322-9, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21323932

RESUMO

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.


Assuntos
Anticorpos Anti-Helmínticos/sangue , Neurocisticercose/diagnóstico , Neurocisticercose/imunologia , Parasitologia/métodos , Biblioteca de Peptídeos , Peptídeos , Taenia solium/imunologia , Animais , Galinhas , Humanos , Imunoglobulina G/sangue , Peptídeos/isolamento & purificação , Sensibilidade e Especificidade , Testes Sorológicos/métodos , Soro/química
12.
Vet Parasitol ; 158(4): 319-28, 2008 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-18977600

RESUMO

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 5p.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.


Assuntos
Genes MHC da Classe II/genética , Genes MHC Classe I/genética , Enteropatias Parasitárias/imunologia , Pneumopatias Parasitárias/imunologia , Estrongiloidíase/imunologia , Animais , Enteropatias Parasitárias/parasitologia , Enteropatias Parasitárias/patologia , Intestinos/patologia , Pulmão/patologia , Pneumopatias Parasitárias/parasitologia , Pneumopatias Parasitárias/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Strongyloides , Estrongiloidíase/parasitologia , Estrongiloidíase/patologia
13.
Vet Parasitol ; 147(1-2): 132-9, 2007 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-17448603

RESUMO

Strongyloides stercoralis is an intestinal nematode with worldwide distribution, particularly in tropical and subtropical regions. Due to the low sensitivity of traditional parasitological methods, the detection of serum specific antibodies may serve as an alternative test for the diagnosis. The aims of the present study were to verify the occurrence of S. stercoralis and the presence of specific IgG antibodies to the parasite in kennel dogs and keepers, using parasitological and serological assays. A total of 181 dogs were examined from 7 breeding kennels in the city of Uberlândia, southeastern region of Brazil and distributed as follows: kennel A (n=41), kennel B (n=16), kennel C (n=11), kennel D (n=63), kennel E (n=11), kennel F (n=18) and kennel G (n=21). Fecal and serum samples from 11 keepers responsible for kennel cleaning and dog control were also collected in five of the seven kennels (two from kennel A, one from kennel B, four from kennel D, two from kennel E and two from kennel G). Overall, enteroparasites were detected by parasitological assays in 66, 36.5% (95% CI: 2.5-43.4%) of the 181 dogs tested. Only one (0.6%) dog was copropositive for S. stercoralis. Among the keepers only one fecal sample, 9.1% (95% CI: 8.6-9.4%) was positive for hookworm by the Lutz method. Serological assays showed that 44 (24.3%) of the 181 dogs were seropositive for S. stercoralis in at least one of the tests in the following kennels: 21 (11.6%) in kennel A; 1 (0.6%) in kennel B; 5 (2.7%) in kennel C; 6 (3.3%) in kennel D; 1 (0.6%) in kennel E; 9 (4.9%) in kennel F and 1 (0.6%) in kennel G. Among the keepers no S. stercoralis seropositive samples were identified using IFAT but 2 (18.2%) of the keepers from kennel D and 1 (9.1%) from kennel G were seropositive by ELISA. The present study demonstrated that the occurrence of S. stercoralis infection in kennel dogs and keepers is low in the city of Uberlândia and that serological assays can contribute to the diagnosis of canine as well as human strongyloidiasis.


Assuntos
Anticorpos Anti-Helmínticos/sangue , Doenças do Cão/epidemiologia , Strongyloides stercoralis/isolamento & purificação , Estrongiloidíase/epidemiologia , Animais , Brasil/epidemiologia , Doenças do Cão/diagnóstico , Cães , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoglobulina G/sangue , Masculino , Prevalência , Estrongiloidíase/diagnóstico
14.
Vet Parasitol ; 136(2): 137-45, 2006 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-16321470

RESUMO

Canine strongyloidiasis is a parasitic infection caused by the nematode Strongyloides stercoralis and presents a great zoonotic potential. Its confirmation, using coproparasitological methods, is difficult. The detection of serum specific antibodies, however, may facilitate the diagnosis. The aims of this study were to determine the presence of S. stercoralis through the use of parasitological methods and to detect specific antibodies to the parasite in serum samples from domestic dogs by using the indirect fluorescent antibody test (IFAT) on slides and the enzyme-linked immunosorbent assay (ELISA). A total of 215 dogs of various breeds, from the cities of Uberlândia, Araxá and Campo Belo in the State of Minas Gerais, were examined and distributed according to age into the following groups: (I) 19 males and 20 females of 1-2 months old; (II) 11 males and 20 females of 2-month- to 1-year-old and (III) 41 males and 104 females, from 1 to 7 years old. Coproparasitological results showed that 63/215 (29.3%) of the dogs presented some kind of parasite, with two (0.9%) dogs (one from Araxá and the other from Uberlândia) passing S. stercoralis larvae in the feces. Serological results revealed antibodies to S. stercoralis in 45/215 (20.9%) of the dogs, with seropositivity rates of 0% (0/39) in Group I, 22.6% (7/31) in Group II, and 26.2% (38/145) in Group III. No serological cross-reactivity between S. stercoralis and hookworms or Ascaridae was found. Hookworm infections were seen in 31 dogs, but only one of these dogs (infected with both hookworm and Cystoisospora spp.) was S. stercoralis seropositive by IFAT. The present study demonstrated, for the first time, natural S. stercoralis infections in dogs diagnosed by coproparasitological and serological methods. It was concluded that the detection of specific antibodies to S. stercoralis by IFAT and ELISA may contribute to the diagnosis of canine strongyloidiasis.


Assuntos
Doenças do Cão/diagnóstico , Ensaio de Imunoadsorção Enzimática/veterinária , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Strongyloides stercoralis/isolamento & purificação , Estrongiloidíase/veterinária , Fatores Etários , Animais , Anticorpos Anti-Helmínticos/sangue , Brasil , Doenças do Cão/parasitologia , Cães , Ensaio de Imunoadsorção Enzimática/métodos , Fezes/parasitologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo/métodos , Humanos , Imunoglobulina G/sangue , Masculino , Strongyloides stercoralis/imunologia , Estrongiloidíase/diagnóstico , Estrongiloidíase/parasitologia
15.
Rev. méd. Chile ; 130(12): 1358-1364, dic. 2002.
Artigo em Espanhol | LILACS | ID: lil-356137

RESUMO

BACKGROUND: Strongyloides stercoralis is a world wide distributed small intestinal nematode parasite. In immunocompetent individuals S stercoralis can produce asymptomatic infections or a moderate clinical picture of diarrhea, some cases become chronic. In immunocompromised patients, a disseminated disease may appear, sometimes fatal. In Chile, there is little epidemiological information about S stercoralis infections and appropriate diagnostic techniques are usually not used. AIM: To evaluate the yield of an ELISA test for the diagnosis of strongyloidiasis in Chilean patients. MATERIAL AND METHODS: Ten serum samples from patients with S stercoralis infections confirmed by a positive stool examination, 66 samples from individuals with other infections by tissue helminthes (24 toxocariasis, 15 trichinellosis, 11 hydatidosis, 12 fascioliasis and 4 cysticercosis), 13 samples from subjects with autoimmune diseases and 49 samples from apparently healthy individuals with a normal eosinophil count, were studied. ELISA antigen was prepared using a filariform larval extract obtained from a murine species of Strongyloides, maintained in laboratory animals. RESULTS: Using 0.33 optical density units as a cut off value, 9 of 10 sera of S stercoralis infected individuals, had a positive ELISA test. No cross reactions were observed with sera of patients with other helminthic infections, autoimmune diseases or in healthy individuals. Thus, specificity, positive and negative predictive values were 100 per cent. CONCLUSIONS: The results obtained are similar with those found by other investigators. ELISA test for strongyloidiasis is a useful tool for the diagnosis of clinical cases and for seroepidemiological studies of this nematode infection in Chile.


Assuntos
Humanos , Criança , Adulto , Estrongiloidíase/diagnóstico , Strongyloides stercoralis/imunologia , Doenças Autoimunes , Ensaio de Imunoadsorção Enzimática , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/sangue , Chile , Estrongiloidíase/sangue , Estrongiloidíase/imunologia , Helmintíase/sangue , Helmintíase/diagnóstico , Helmintíase/imunologia , Sensibilidade e Especificidade
17.
Trop Med Int Health ; 6(3): 219-26, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11299039

RESUMO

We compared saline (S) and sodium dodecyl sulphate (SDS) extracts from Taenia solium (homologous species - HO) and Taenia crassiceps (heterologous species - HE) metacestodes in order to detect IgG by ELISA and immunoblot assay (IBA) in cerebrospinal fluid (CSF) for the diagnosis of human neurocysticercosis (NC). CSF samples were obtained from 93 patients. Of these, 40 had NC, five had a diagnosis of probable NC, nine had central nervous system schistosomiasis or strongyloidiasis and 39 had other neurological alterations. Samples were analysed by ELISA and the results were compared with IBA in all samples with confirmed and probable NC diagnosis, in all samples with other central nervous system parasitic infection, and in 10 of those with another neurological alterations. ELISA sensitivity was 100%, 85%, 95% and 87.5% for the S-HO, S-HE, SDS-HO and SDS-HE extracts, respectively, and ELISA specificity was 100% for S-HO, S-HE, SDS-HO extracts and 97.9% for SDS-HE antigen. Immunodominant peptides detected by IBA were, by decreasing percentage of recognition: 64-68 and 45 kDa for S-HO; 108-114, 92-95, 64-68, 83 and 88 kDa for S-HE; 64-68, 108-114, 77 and 86 kDa for SDS-HO; and 108-114, 88 and 92-95 kDa for SDS-HE. Overall the homologous antigenic extracts showed higher sensitivity than the heterologous extracts in the diagnosis of NC in CSF samples. The heterologous extracts contained most of the immunodominant peptides presented in the homologous extracts, which are recognized by IgG antibodies in CSF samples.


Assuntos
Anticorpos Anti-Helmínticos/líquido cefalorraquidiano , Imunoglobulina G/líquido cefalorraquidiano , Neurocisticercose/diagnóstico , Taenia/imunologia , Animais , Ensaio de Imunoadsorção Enzimática , Humanos , Immunoblotting , Peso Molecular , Testes Sorológicos , Cloreto de Sódio/farmacologia , Dodecilsulfato de Sódio/farmacologia
18.
Trop Med Int Health ; 5(6): 443-9, 2000 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10929145

RESUMO

A comparative study of total saline extract (SE) and cyst vesicular fluid (VF) of Taenia solium metacestodes by ELISA and Western blotting assay (WB) tests was conducted to detect IgG in sera for diagnosis of human cysticercosis. Sera were obtained and analysed by ELISA in 1 : 20 and 1 : 100 dilutions from 208 individuals: 22 confirmed neurocysticercosis (NC) (group 1), 101 suspected NC (group 2), 55 with various intestinal parasitosis (group 3) and 30 healthy individuals (group 4). The WB test was carried out on SE and VF extracts with and without reducing agent, 2-beta-mercaptoethanol (2-ME) in 20 sera of each group. WB using extracts without 2-ME and ELISA at 1 : 100 dilution were compared in 20 sera from each group; sensitivity and specificity were calculated using samples from groups 1, 3 and 4. By ELISA, in the 1 : 100 sera dilution reactivity was reduced for both antigens without changes in the sensitivity of the test. By WB, antigens treated with 2-ME demonstrated low specificity. For SE and VF antigens, the proteins of 24, 39-42, 47-52, 56, 64-68, 126-155 kDa and 18, 24, 26-28, 32-36, 47-52, 75 kDa, respectively, were considered immunodominant markers, with high indices of specificity, suggesting a profile for NC patients. However, as the sensitivity was found to be low, it might still not be a definitive test for NC when used alone. These data suggest WB as an indicative test to determine exposure to T. solium. ELISA and WB together may supply reliable results for the diagnosis of human cysticercosis, since appropriate purified antigens are not available yet.


Assuntos
Anticorpos Anti-Helmínticos/sangue , Imunoglobulina G/sangue , Neurocisticercose/diagnóstico , Taenia/imunologia , Adolescente , Adulto , Idoso , Animais , Western Blotting/normas , Criança , Pré-Escolar , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática/normas , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Taenia/isolamento & purificação
19.
Rev Inst Med Trop Sao Paulo ; 42(1): 51-5, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-10742728

RESUMO

Parasitological and immunological diagnoses were part of a study conducted among 151 children, 83 immunocompromised (IC) and 68 non-immunocompromised (non-IC) aged from zero to 12, seen at the University Hospital, Universidade Federal de Uberlândia, State of Minas Gerais, Brazil, from February, 1996, to June, 1998. Three fecal samples from each child were analyzed for the parasitological diagnosis by Baermann-Moraes and Lutz methods. The immunological diagnosis to detect IgG and IgM antibodies was carried out by the indirect immunofluorescence antibody test (IFAT) with cryo-microtome sections of Strongyloides stercoralis and Strongyloides ratti larvae as antigens and by the ELISA test with an alkaline extract of S. ratti as the antigens. Of the 151 children 5 (3.31%) were infected with larvae of S. stercoralis (2 cases IC, 2.41%, and 3 cases non-IC, 4.41%). The IFAT-IgG detected 7 (8.43%) serum samples positive among IC, and 2 (2.94%) cases among non-IC. The ELISA-IgG test detected 10 (12.05%) serum samples positive among IC, and 1 (1.47%) case among non-IC. The IFAT-IgM detected 6 (7.22%) positive cases among IC, and 3 (4.41%) cases among non-IC. ELISA-IgM test detected 10 (12.05%) positive cases among IC, and 3 (4.41%) cases among non-IC. It was concluded that the immunological tests can help in the diagnosis of strongyloidiasis in immunocompromised children.


Assuntos
Anticorpos Anti-Helmínticos/sangue , Hospedeiro Imunocomprometido , Strongyloides stercoralis/isolamento & purificação , Estrongiloidíase/diagnóstico , Animais , Brasil , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Lactente , Recém-Nascido , Strongyloides stercoralis/imunologia , Strongyloides stercoralis/parasitologia , Estrongiloidíase/sangue
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