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Concordancia entre ELISA e IFI para la determinación de anticuerpos tipo IgG contra Toxoplasma gondii Concordance between ELISA and IFA, for IgG antibodies detection against Toxoplasma gondii  [cached]
Liliana Jazmín Cortés,Lorena Mancera
Infectio , 2009,
Abstract: Objetivo. Medir el grado de concordancia entre las técnicas de inmunofluorescencia indirecta y de ELISA, empleadas para la detección de anticuerpos IgG anti-Toxoplasma gondii en humanos. Materiales y métodos. Se llevó a cabo un estudio descriptivo comparativo de dos técnicas diagnósticas para la detección de anticuerpos IgG anti-T. gondii, en 243 sueros humanos del banco de muestras del Laboratorio de Parasitología-Red Nacional de Laboratorios del Instituto Nacional de Salud, recolectados durante los a os 2000 a 2006. Resultados. El porcentaje de reactividad para la detección de anticuerpos IgG anti-T. gondii fue de 54% (131 sueros) por la técnica IFI y de 56,3% (137 sueros) por la técnica ELISA; además, el 46% (112 sueros) de las muestras fueron negativas por la técnica IFI y el 44% (106 sueros) negativas fueron por la técnica ELISA. El valor del índice kappa fue de 0,916 (IC95% 0,866-0,976), el cual refleja una concordancia casi absoluta entre las dos técnicas. Conclusiones. El índice kappa muestra una concordancia casi absoluta entre las técnicas, IFI y ELISA, lo cual hace que los laboratorios departamentales de salud pública del país puedan cumplir con la determinación de anticuerpos IgG anti-T. gondii como prueba diagnóstica en el control prenatal, cuando no se cuente con la infraestructura necesaria para la técnica de IFI, teniendo en cuenta que la de ELISA está fundamentada en el grado de afinidad de anticuerpos por un antígeno determinado y no mide el nivel real de éstos en una muestra de suero, como sí lo hace la técnica de IFI.
Concordancia entre ELISA e IFI para la determinación de anticuerpos tipo IgG contra Toxoplasma gondii
Cortés,Liliana Jazmín; Mancera,Lorena;
Infectio , 2009,
Abstract: objetivo. medir el grado de concordancia entre las técnicas de inmunofluorescencia indirecta y de elisa, empleadas para la detección de anticuerpos igg anti-toxoplasma gondii en humanos. materiales y métodos. se llevó a cabo un estudio descriptivo comparativo de dos técnicas diagnósticas para la detección de anticuerpos igg anti-t. gondii, en 243 sueros humanos del banco de muestras del laboratorio de parasitología-red nacional de laboratorios del instituto nacional de salud, recolectados durante los a?os 2000 a 2006. resultados. el porcentaje de reactividad para la detección de anticuerpos igg anti-t. gondii fue de 54% (131 sueros) por la técnica ifi y de 56,3% (137 sueros) por la técnica elisa; además, el 46% (112 sueros) de las muestras fueron negativas por la técnica ifi y el 44% (106 sueros) negativas fueron por la técnica elisa. el valor del índice kappa fue de 0,916 (ic95% 0,866-0,976), el cual refleja una concordancia casi absoluta entre las dos técnicas. conclusiones. el índice kappa muestra una concordancia casi absoluta entre las técnicas, ifi y elisa, lo cual hace que los laboratorios departamentales de salud pública del país puedan cumplir con la determinación de anticuerpos igg anti-t. gondii como prueba diagnóstica en el control prenatal, cuando no se cuente con la infraestructura necesaria para la técnica de ifi, teniendo en cuenta que la de elisa está fundamentada en el grado de afinidad de anticuerpos por un antígeno determinado y no mide el nivel real de éstos en una muestra de suero, como sí lo hace la técnica de ifi.
Aplicación de herramientas serológicas y moleculares para el diagnóstico de coriorretinitis por Toxoplasma gondii Application of serological and molecular tools for diagnosis of chorioretinitis caused by Toxoplasma gondii  [cached]
Bernardo Regalado Andújar,Martha Solangel Rodríguez Pe?a,Jorge Fraga Nodarse,Lázara Rojas Rivero
Revista Cubana de Medicina Tropical , 2013,
Abstract: Introducción: Toxoplasma gondii, agente causal de la toxoplasmosis, es un protozoario intracelular obligado que puede afectar al globo ocular, siendo la causa más común de uveítis posterior. Objetivo: determinar la utilidad de las técnicas serológicas y moleculares para el diagnóstico de la toxoplasmosis ocular en pacientes con uveítis. Métodos: se dise ó un estudio de corte transversal para comparar un grupo de pacientes afectados por coriorretinitis toxoplásmica y otro con signos sugestivos de coriorretinitis no toxoplásmica. En ambos grupos se utilizaron métodos serológicos como la inmunofluorescencia indirecta y la prueba de inmunoensayo enzimático (ELISA), y la reacción en cadena de la polimerasa para el diagnóstico de la toxoplasmosis ocular. Resultados: las técnicas serológicas permitieron detectar los anticuerpos IgG anti-Toxoplasma en 100 % de los pacientes con coriorretinitis toxoplásmica, de ellos fueron positivos a los anticuerpos IgM solo 3 pacientes y 1 tuvo un resultado débil a la avidez IgG. La reacción en cadena de la polimerasa detectó el ADN de Toxoplasma gondii en 15 de los 47 pacientes para una sensibilidad de 31,9 %. Conclusiones: se reporta la detección de anticuerpos anti-Toxoplasma, en todos los casos de coriorretinitis toxoplásmica a través de los métodos serológicos, aunque la técnica más sensible fue la inmunofluorescencia indirecta. Se empleó la detección molecular del ADN de Toxoplasma gondii en pacientes con toxoplasmosis ocular, por primera vez en Cuba. Los resultados permiten sugerir el uso de técnicas serológicas y moleculares que ayuden a confirmar el diagnóstico de infección por Toxoplasma en pacientes con coriorretinitis. Introduction: Toxoplasma gondii, the causative agent of toxoplasmosis, is an intracellular protozoan that can affect the eye, and be the most common cause of posterior uveitis. Objective: to determine the usefulness of serological and molecular techniques for the diagnosis of ocular toxoplasmosis in patients with uveitis. Methods: a cross-sectional study was designed in order to compare a group of patients with toxoplasmic retinochoroiditis and another affected by non-toxoplasmic retinochoroiditis. In both groups serological methods such as indirect immunofluorescence assay, immunoassay (ELISA), and the polymerase chain reaction were used for the diagnosis of ocular toxoplasmosis. Results: serological techniques allowed detecting anti-Toxoplasma IgG antibodies in 100 % of patients with toxoplasmic retinochoroiditis, and only 3 patients were positive to IgM antibodies. Finally, only one had a weak re
Analysis Concentration of Toxoplasma gondii on Anti-Toxoplasma IgG-IgM Antibody Levels, and the Outcomes of Pregnancy in Mice Balb/c  [PDF]
Tigor Peniel Simanjuntak, Mochammad Hatta, Robert H. Sirait, Marni Br Karo, Lenny Irmawaty Sirait, Tetty Rina Aritonang, Syahrul Rauf, Ressy Dwiyanti
Open Journal of Obstetrics and Gynecology (OJOG) , 2017, DOI: 10.4236/ojog.2017.73030
Abstract: Introduction: Toxoplasma gondii is an obligate intracellular protozoan parasite that can infect any warm blood vertebrae, and if first trimester pregnant woman infected, it may cause abortion. The objective is to prove the effect of the Toxoplasma gondii concentration in anti-toxoplasma IgG-IgM antibody levels, and the outcomes of Balb/c mice pregnancies. Materials and Methods: The study was conducted in Balb/c mice with inclusion criteria, and was conditioned pregnant. The pathogen strains of Toxoplasma gondii tachyzoite injected intraperitoneally. The blood samples were taken serially to be tested for anti-toxoplasma IgG-IgM antibody levels. After the mice were injected with tachyzoite, they are assessed every day to observe their body weight, vaginal bleeding, and labor. Anti-toxoplasma IgG-IgM antibody levels examined using qualitative mouse IgG-IgM antibody ELISA KIT. Results: Anti-toxoplasma IgM antibody levels increased significantly after 24 hours of injection tachyzoites in all dose groups, and remained high through day 21. Anti-toxoplasma antibody IgG levels increased significantly after 72 hours post injection and remained elevated until day 21. The incidence of abortion is 100% in mice which injected tachyzoite levels 1 × 103 and 1 × 104, and the incidence of abortion approximately 2 - 4 days post injection. 100% of mice that were injected with tachyzoites 1 × 101 and 1 × 102 have labor at term. Physical anomaly was found in baby mice from mice that were injected with tachyzoite 1 × 102. Conclusion: There is a significant correlation between the concentrations of Toxoplasma gondii tachyzoite with anti-toxoplasma IgG-IgM antibody levels, and there is a significant relationship between the concentrations of tachyzoite with abortion.
Determination of Antibodies (IgG, IgM) against Toxoplasma gondii in Patients with Cancer
M Ghasemian,Sh Maraghi,J Saki,M Pedram
Iranian Journal of Parasitology , 2007,
Abstract: Background: The aim of this study was determination of antibodies (IgG, IgM) against Toxoplasma in malignant patients in order to refer the patients on time to the physician for treatment.Methods: This study was carried out on 252 malignant patients and 252 healthy normal subjects (as control) obtained from Shafa Hospital and Medical Diagnostic Laboratory (Iran-Zamin), in Ahwaz city. Patient's information was recorded in a questionnaire before sampling. Serum samples of patients were examined for IgG and IgM antibodies by ELISA technique using Trinity kits. Results: The results of this study revealed the presence of Toxoplasma antibodies in 114 (45.2%) cases of patients who were positive for Toxoplasma IgG antibodies, and 26 (10.3%) cases were confirmed to be positive for Toxoplasma IgM antibodies and also 17 (6.7%) of cases had both IgG and IgM antibodies against Toxoplasma gondii. In control group 92 (36.5%) cases and 15 (6%) cases revealed seropositive for IgG and IgM antibodies, respectively. There were no significant differences between sex, close contact with cat, living region, chemotherapy, and seropositivity rate of toxoplasmosis in patients. Comparing the age groups, the highest seropositive rate showed in the age of 51 years or higher, and their rates had tendency to increase with age in both groups. No seropositivity significant relationship was found between patients and control group.Conclusion: According to the prevalence of positive cases in these patients, it is necessary to examine the patients for toxoplasmosis before, during and after chemotherapy.
Toxoplasma gondii IgG antibodies in HIV/AIDS patients attending hospitals in Makurdi metropolis, Benue state, Nigeria  [PDF]
Amuta E.U,Amali O,Jacob S.E,Houmsou R.S
International Journal of Medicine and Biomedical Research , 2012,
Abstract: Background: Toxoplasmosis is caused by Toxoplasma gondii, a parasite that gradually evolved to be the most opportunistic parasite that complicates the course of HIV/AIDS in developing countries. Aim: This study was undertaken to investigate the presence of Toxoplasma gondii IgG antibodies in HIV-infected patients attending hospitals in Makurdi metropolis, Benue State, Nigeria. Materials and Methods: The Enzyme-linked immunosorbent assay (ELISA) technique was used to determine the presence of Toxo-IgG antibodies in blood samples collected from the HIV/AIDS patients and their CD4 counts were estimated using flow cytometry. Questionnaires were also administered to obtain information on their socio-demographic status. Results: Thirty-nine, 39 (10.8%) were screened positive for Toxo-IgG antibodies out of the 360 HIV/AIDS patients enrolled. Males (10.3%) and females (11.2%) had similar seroprevalence of Toxo-IgG with no significant difference (χ2= 0.001, p>0.05). The presence of Toxoplasma IgG antibodies was found to be highest in the ≥ 54 years age group. A significant difference was observed in the seroprevalence of Toxoplasma IgG among age groups (χ2=11.56, p<0.05). Females with CD4 T-cell count ≤ 200 cells/mm3 recorded higher seroprevalence (73.7%) of Toxoplasma IgG. There was no significant difference in the seroprevalence of Toxoplasma IgG in relation to CD4 T-cell counts (χ2= 2.3, p > 0.05). Conclusion: Further investigations are still needed to clarify the exact relationship of the parasite infection, its effects on the HIV-infected and uninfected individuals and the detection of their major means of transmission.
Detection of anti-Toxoplasma gondii antibodies in experimentally and naturally infected non-human primates by Indirect Fluorescence Assay (IFA) and indirect ELISA
Bouer, Andréa;Werther, Karin;Machado, Rosangela Zacarias;Nakaghi, Andréa Cristina Higa;Epiphanio, Sabrina;Cat?o-Dias, José Luiz;
Revista Brasileira de Parasitologia Veterinária , 2010, DOI: 10.4322/rbpv.01901005
Abstract: the indirect fluorescence assay (ifa) and the indirect elisa were comparatively used to detect igg and igm antibodies for toxoplasma gondii in experimentally and naturally infected primates. in the experimentally infected group, antibodies of diagnostic value were detected at day 9 post-infection (pi) with the ifa (igg and igm) and with igg-elisa. igm-elisa detected antibodies for t. gondii starting at day 3 pi until the end of the experiment (102 days pi). of the 209 naturally infected sera tested, from many zoos of state of sao paulo, 64.59 and 67.94% were positive in the igg-ifa test and igg-elisa respectively. igm-elisa test detected seropositivity in 52.63% of the sera although igm-ifa test detected it in only in 0.96% of the samples. the differential toxoplasmosis diagnosis was accomplished with neospora caninum by ifa, observing 61 (29.2%) seropositive animals for this parasite and 149 (70.8%) negative. sixty animals were positive for both t. gondii and n. caninum. pneumonia, splenomegaly, and intestinal ulcers were macroscopically observed. unremarkable interstitial pneumonia, enteritis, colitis, splenitis, and glomerulitis were microscopically observed. the immunohistochemical stain could not detect the presence of t. gondii in the tissues of the animals infected experimentally.
Designing of ELISA kit for diagnosis of anti-Toxoplasma specific IgG, using exerted/secreted antigens and its
S. H. Abdollahi, Ph.D.,M. Kazemi, Ph.D.
Journal of Mazandaran University of Medical Sciences , 2008,
Abstract: Background and Purpose: Many studies report that Toxoplasma gondii excreted / secreted antigens (E/SA) appear to be a suitable marker for toxoplasmosis serodiagnosis. Most of these studies have used E/SA, obtained from supernatant of Toxoplasma cell culture, or by incubating tachyzoites in cell free media (RPMI-1640). The present study for the detection of Toxoplasma IgG in human serum was evaluated, using the components of peritoneal fluid of infected mice (as another source of E/SA). Materials and Methods: Peritoneal fluids of mice infected by interaperitoneal (IP) inoculation of Toxoplasma tachyzoites were collected after 3 days and centrifuged at 750×g for 15 minutes. Then the supernatant was precipitated with ammonium sulphate solution (40% saturated) and used as components encompassing E/SA.Forty (40) none- infected ( without anti- toxoplasma antibodies) and thirty- two (32) positive (with IgG to toxoplasma) human serum samples were selected (all sera were first tested by standard method for detection of IgG antibodies anti-T. gondii) then, the sera were tested by ELISA using E/SA.Results: The cut-off point with 95% confidences was found to be 0.78. Moreover, sensitivity and specificity of the method were determined to be 84% and 92%, respectively.Conclusion: The present results indicate that peritoneal exudates from mice infected with T.gondii, may be used as a source of anti-genic material for the detection of Toxoplasma-specific IgG. Furthermore, it may be valuable for the development of new tools in the serodiagnosis of toxoplasmosi
Perbedaan Metode ELISA Sandwich A dan B dalam Deteksi Antigen Membran Toxoplasma gondii  [PDF]
MURKATI,NOERHAYATI SURIPTO,,SUPARGIYONO,,S. TJOKROSONTO
Bioteknologi , 2004,
Abstract: Spreading of toxoplasmosis to fetus can by placenta, so it caused theabortion, born dead or congenital defect. To diagnosis this disease for fixed the acute infection must get the significant increasing of IgG by the soft fee. The objections of this study are to know the difference between ELISA Sandwich A and B in detecting of membrane antigen of Toxoplasma gondii (T. gondii) in placenta tissue of pregnant women three-semester I and II with spontaneous abortion in Surakarta. One hundred serum and placenta tissue samples of pregnant women three-semester I and II with spontaneous abortion are got from dr. Muwardi Hospital. IgM anti Toxo from serum was examined by Toxo ISAGA Kit and IgG anti Toxo by Toxo Screen DA Kit. Detecting of membrane antigen of T. goodie from placenta tissue were done by ELISA Sandwich A and B. The result of this experiment showed that 33% were positive IBM and or Gig anti Toxo. Detection of membrane antigen toward 33 samples with positive Toxo (IgG positive) was highly significant different between ELISA Sandwich A (3% positive) toward ELISA Sandwich B (72.7% positive).
Early diagnosis of congenital toxoplasmosis in newborn infants using IgG subclasses against two Toxoplasma gondii recombinant proteins
Silva, Carlos Henryque de Souza e;Andrade, Gláucia Queiroz de;Januário, José Nélio;Carneiro, Ana Carolina de Aguiar Vasconcelos;Carneiro, Mariangela;Vasconcelos-Santos, Daniel Vitor;Vitor, Ricardo Wagner de Almeida;
Memórias do Instituto Oswaldo Cruz , 2012, DOI: 10.1590/S0074-02762012000300008
Abstract: the aim of this work was to evaluate the utility of elisa-based testing of total igg (iggt) antibodies and its subclasses (igg1, igg2, igg3 and igg4) against soluble (stag) and recombinant (rsag1 and rmic3) antigens of toxoplasma gondii for diagnosing congenital toxoplasmosis. sera from 217 newborns initially testing positive for specific igm in filter paper dried blood spots were tested for specific igm and igg by elfa-vidas?. congenital toxoplasmosis was confirmed in 175 and ruled out in 42 infants. the validity of the elisa tests was determined using the persistence of igg antibodies (elfa-vidas? kit) at the end of 12 months, which is considered the reference test for the diagnosis of congenital toxoplasmosis. the frequency of positivity with iggt against stag, rsag1 and rmic3 was found in 97.2%, 96.3% and 80.2%, respectively, of the newborns with confirmed congenital toxoplasmosis. igg1 reacted with all three antigens, while igg3 and igg4 reacted preferentially with rmic3. higher mean values of reactivity (sample optical density/cut-off) were found for all subclasses when using rmic3. all of the antigens showed high sensitivity and low specificity in detecting anti-t. gondii iggt and igg1 and low sensitivity and high specificity in detecting igg3 and igg4. in conclusion, the combined detection of igg antibody subclasses against recombinant toxoplasmic antigens may be useful for the early diagnosis of congenital toxoplasmosis.
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