scholarly journals AN EVALUATION STUDY OF ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) USING RECOMBINANT PROTEIN GRA1 FOR DETECTION OF IgG ANTIBODIES AGAINTS TOXOPLASMA GONDII INFECTIONS

2017 ◽  
Vol 6 (5) ◽  
pp. 105
Author(s):  
Nina Difla Muflikhah ◽  
Wayan Tunas Artama
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Protein ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Low Cost ◽  
Evaluation Study ◽  
Time Frame ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibody ◽  
Elisa Kit

Toxoplasmosis is an infectious disease caused by Toxoplasma gondii, an intracellular protozoan parasite that live inside the cells of the reticulo endothelial and parenchymal cells of human and animals (mammals and birds). Some cases of toxoplasmosis usually have no symptoms, but in any cases caused severe symptoms, such as hydrocephalus, microcephalus, intracranial calcification, retinal damage, brain abscess, mental retardation, lymphadenopathy, and others. Its severe symptoms usually showed a long time after first exposure, except symptoms showed by congenital transmission caused by infected mother. Early diagnosis is important to prevent the illness but methods for toxoplasmosis screening are still too expensive for developing country. Enzyme-linked immunosorbent assay (ELISA) allow the testing of a large number samples within short time frame and based on antibody or antigen detection. This study aimed to know the sensitivity and specificity of recombinat protein GRA1 as antigen using ELISA methods. We tested the sensitivity and spesificity of GRA1 protein as antigen in ELISA methods to diagnose toxoplasmosis and compared with ELISA Kit Commercial. Reliable laboratory testing is important to detect Toxoplasma gondii infection, and focused to improving the low cost and easy-to-use diagnostic instrument. Seventy sera collected and tested using both indirect ELISA, commercial ELISA kit and GRA1 protein coated as antigen. Fourty eight and fifty one samples showed positive IgG antibody result of ELISA-GRA1 and ELISA kit. Negative sample tested by ELISA-GRA1 was 22 samples and 19 sample tested by ELISA Kit. The sensitivity and specificity of GRA1-based on ELISA were 100% and 86.36%, positive prediction value (ppv) was 94.11%. These data indicate that the recombinant protein GRA1 is a highly immunogenic protein in human toxoplasmosis and become a promising marker for the screening of toxoplasmosis.

Analysis of the antibodies anti-Toxoplasma gondii by ELISA based on two diagnostic antigens: rSAG1 and rBAG1

2011 ◽  
Vol 56 (4) ◽  
Author(s):  
Min Liu ◽  
Tao Wang ◽  
Hua Li ◽  
Ju-Ying Li ◽  
Han Zhong ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Immunosorbent Assay ◽  
Blood Donors ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibody ◽  
Neuropsychiatric Disease ◽  
Increased Risk ◽  
Positive Rate ◽  
Toxoplasma Gondii Infection ◽  
Uncertain Etiology

AbstractSchizophrenia is a serious neuropsychiatric disease of uncertain etiology. Previous studies have demonstrated that antibodies to Toxoplasma gondii infection are associated with an increased risk of schizophrenia. The objective of this study was to analyze anti-T. gondii antibodies in 477 Chinese schizophrenia patients using an enzyme-linked immunosorbent assay (ELISA) based on recombinant surface antigen 1 (rSAG1), recombinant bradyzoite antigen 1 (rBAG1) and the soluble tachyzoite antigens (STAg) of T. gondii RH strain. Results showed that among the sero-positives (IgG and/or IgM) for T. gondii infection examined in schizophrenia patients, sero-positive samples for rSAG1, rBAG1 and STAg were 20.5% (98/477), 20.5% (98/477) and 23.5% (112/477) respectively, while compared to 210 blood donors, sero-positive (IgG and/or IgM) samples for these antigens (rSAG1, rBAG1 and STAg) were only 5.7% (12/210), 6.2% (13/210) and 5.7% (12/210), respectively. Furthermore, when IgG antibody reaction in the schizophrenia sera was compared with the rBAG1 and rSAG1, results demonstrated that beside the cases which can be detected by both rSAG1 and rBAG1, some sero-positive for T. gondii in schizophrenia sera can only be detected either by rSAG1 or rBAG1. This phenomenon was also observed in the detection of IgM with rSAG1 and rBAG1. 5.9% (28/477) of cases of schizophrenia which are positive for IgG or IgM by rSAG1 are negative for STAg, while 9.2% (44/477) of the schizophrenia cases which are positive for IgG or IgM by rBAG1 are negative for STAg. Although STAg can also be used to diagnose T. gondii infection from schizophrenia patients, it may not actually indicate the infection as some positive samples may be mistakenly considered to be negative. In conclusion, our results demonstrate that the sero-positive rate for T. gondii in the Chinese schizophrenia patients was higher than blood donors. More importantly, our results provide evidence that the combination of rSAG1 and rBAG1 antigens in the diagnosis of T. gondii infection could closely reflect the actual infection of this parasite in schizophrenia patients.

scholarly journals Development and evaluation of recombinant GRA8 protein for the serodiagnosis of Toxoplasma gondii infection in goats

2020 ◽  
Author(s):  
Charoonluk Jirapattharasate ◽  
Ruenruetai Udonsom ◽  
Apichai Prachasuphap ◽  
Kodcharad Jongpitisub ◽  
Panadda Dhepakson
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Protein ◽  
Sensitivity And Specificity ◽  
Western Blotting ◽  
Sodium Dodecyl ◽  
Laboratory Animals ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Field Samples ◽  
Specific Igg

Abstract Background The development of sensitive and specific methods for detecting Toxoplasma gondii infection is critical for preventing and controlling toxoplasmosis in humans and other animals. Recently, various recombinant proteins have been used in serological tests for diagnosing toxoplasmosis. The production of these antigens is associated with live tachyzoites obtained from cell cultures or laboratory animals for genomic extraction to amplify target genes. Synthetic genes have gained a key role in recombinant protein production. For the first time, we demonstrated the production of the recombinant protein of the T. gondii dense granular antigen 8 (TgGRA8) gene based on commercial gene synthesis. Recombinant TgGRA8 plasmids were successfully expressed in an Escherichia coli system. The recombinant protein was affinity-purified and characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Furthermore, the diagnostic potential of the recombinant protein was assessed using 306 field serum samples from goats via indirect enzyme-linked immunosorbent assay (iELISA) and the latex agglutination test (LAT).Results Western blotting using known positive serum samples from goats identified a single antigen at the expected molecular weight of TgGRA8 (27 kDa). iELISA illustrated that 15.40% of goat samples were positive for T. gondii-specific IgG antibodies. In addition, TgGRA8 provided high sensitivity and specificity, with significant concordance (91.83) and kappa values (0.69) compared with the results obtained using LAT.Conclusion Our findings highlight the production of a recombinant protein from a synthetic TgGRA8 gene and the ability to detect T. gondii infection in field samples. The sensitivity and specificity of TgGRA8 demonstrated that this protein could be a good serological marker for detecting specific IgG in goat sera

scholarly journals Development and Evaluation of Recombinant GRA8 Protein for the Serodiagnosis of Toxoplasma Gondii Infection in Goats

2020 ◽  
Author(s):  
Charoonluk Jirapattharasate ◽  
Ruenruetai Udonsom ◽  
Apichai Prachasuphap ◽  
Kodcharad Jongpitisub ◽  
Panadda Dhepakson
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Protein ◽  
Sensitivity And Specificity ◽  
Western Blotting ◽  
Sodium Dodecyl ◽  
Laboratory Animals ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Field Samples ◽  
Specific Igg

Abstract Background The development of sensitive and specific methods for detecting Toxoplasma gondii infection is critical for preventing and controlling toxoplasmosis in humans and other animals. Recently, various recombinant proteins have been used in serological tests for diagnosing toxoplasmosis. The production of these antigens is associated with live tachyzoites obtained from cell cultures or laboratory animals for genomic extraction to amplify target genes. Synthetic genes have gained a key role in recombinant protein production. For the first time, we demonstrated the production of the recombinant protein of the T. gondii dense granular antigen 8 (TgGRA8) gene based on commercial gene synthesis. Recombinant TgGRA8 plasmids were successfully expressed in an Escherichia coli system. The recombinant protein was affinity-purified and characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Furthermore, the diagnostic potential of the recombinant protein was assessed using 306 field serum samples from goats via indirect enzyme-linked immunosorbent assay (iELISA) and the latex agglutination test (LAT).Results Western blotting using known positive serum samples from goats identified a single antigen at the expected molecular weight of TgGRA8 (27 kDa). iELISA illustrated that 15.40% of goat samples were positive for T. gondii-specific IgG antibodies. In addition, TgGRA8 provided high sensitivity and specificity, with significant concordance (91.83) and kappa values (0.69) compared with the results obtained using LAT.Conclusion Our findings highlight the production of a recombinant protein from a synthetic TgGRA8 gene and the ability to detect T. gondii infection in field samples. The sensitivity and specificity of TgGRA8 demonstrated that this protein could be a good serological marker for detecting specific IgG in goat sera.

scholarly journals Evaluation of a NovelAspergillusAntigen Enzyme-Linked Immunosorbent Assay

2019 ◽  
Vol 57 (7) ◽  
Author(s):  
Karl Dichtl ◽  
Ulrich Seybold ◽  
Steffen Ormanns ◽  
Heidi Horns ◽  
Johannes Wagener
Keyword(s):  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Fungal Infections ◽  
Time Frame ◽  
Enzyme Linked Immunosorbent Assay ◽  
The Novel ◽  
Serum Samples ◽  
European Organization ◽  
Content Type ◽  
Patients At Risk

ABSTRACTInvasive aspergillosis (IA) is a life-threatening infection that mainly occurs in immunocompromised patients. Here, we compared the novelAspergillus-specific galactomannoprotein (GP) enzyme-linked immunosorbent assay (ELISA) (Euroimmun Medizinische Labordiagnostika AG) to the established PlateliaAspergillusgalactomannan (GM) ELISA (Bio-Rad Laboratories) for the detection of IA. A total of 267 serum samples from 45 cases of proven and 4 episodes of probable IA (according to European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group [EORTC/MSG] criteria) and 156 sera from patients without evidence of IA were tested. Pearson’s correlation statistics, as well as sensitivity and specificity, were calculated using manufacturer-recommended (GM) or optimized (GP) cutoff levels.Aspergillus fumigatuswas found in 88% of culture-positive infections. When we analyzed all 423 serum samples, GM and GP tests correlated strongly (r = 0.82,P < 0.0001). Among proven IA cases using samples obtained as closely as possible to the day of proven diagnosis, the sensitivity for both tests was 40%. All cases of probable IA (defined by positive GM testing) were also GP positive. Concordant results of the two ELISAs were obtained in 43 of 49 samples (88%). Extending measurements to all sera available in the time frame of 7 days prior to 7 days after the day of proven diagnosis, 47% and 56% of the cases were detected by the GM and GP tests, respectively. Specificity was 99% for GM and 96% for GP testing. For the diagnosis of IA, sensitivity and specificity of the novel GP ELISA are similar to those of the Platelia GM ELISA. The low sensitivities of both tests underline the need for serial testing in patients at risk for IA.

scholarly journals Development and evaluation of recombinant GRA8 protein for the serodiagnosis of Toxoplasma gondii infection in goats

2020 ◽  
Author(s):  
Charoonluk Jirapattharasate ◽  
Ruenruetai Udonsom ◽  
Apichai Prachasuphap ◽  
Kodcharad Jongpitisub ◽  
Panadda Dhepakson
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Protein ◽  
Sensitivity And Specificity ◽  
Western Blotting ◽  
Sodium Dodecyl ◽  
Laboratory Animals ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Field Samples ◽  
Specific Igg

Abstract BackgroundThe development of sensitive and specific methods for detecting Toxoplasma gondii infection is critical for preventing and controlling toxoplasmosis in humans and other animals. Recently, various recombinant proteins have been used in serological tests for diagnosing toxoplasmosis. The production of these antigens is associated with live tachyzoites obtained from cell cultures or laboratory animals for genomic extraction to amplify target genes. Synthetic genes have gained a key role in recombinant protein production. For the first time, we demonstrated the production of the recombinant protein of the T. gondii dense granular antigen 8 (TgGRA8) gene based on commercial gene synthesis. Recombinant TgGRA8 plasmids were successfully expressed in an Escherichia coli system. The recombinant protein was affinity-purified and characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Furthermore, the diagnostic potential of the recombinant protein was assessed using 306 field serum samples from goats via indirect enzyme-linked immunosorbent assay (iELISA) and the latex agglutination test (LAT).ResultsWestern blotting using known positive serum samples from goats identified a single antigen at the expected molecular weight of TgGRA8 (27 kDa). iELISA illustrated that 15.40% of goat samples were positive for T. gondii-specific IgG antibodies. In addition, TgGRA8 provided high sensitivity and specificity, with significant concordance (91.83) and kappa values (0.69) compared with the results obtained using LAT.ConclusionOur findings highlight the production of a recombinant protein from a synthetic TgGRA8 gene and the ability to detect T. gondii infection in field samples. The sensitivity and specificity of TgGRA8 demonstrated that this protein could be a good serological marker for detecting specific IgG in goat sera

scholarly journals Development and evaluation of recombinant GRA8 protein for the serodiagnosis of Toxoplasma gondii infection in goats

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Charoonluk Jirapattharasate ◽  
Ruenruetai Udonsom ◽  
Apichai Prachasuphap ◽  
Kodcharad Jongpitisub ◽  
Panadda Dhepakson
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Protein ◽  
Sensitivity And Specificity ◽  
Western Blotting ◽  
Sodium Dodecyl ◽  
Laboratory Animals ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Field Samples ◽  
Specific Igg

Abstract Background The development of sensitive and specific methods for detecting Toxoplasma gondii infection is critical for preventing and controlling toxoplasmosis in humans and other animals. Recently, various recombinant proteins have been used in serological tests for diagnosing toxoplasmosis. The production of these antigens is associated with live tachyzoites obtained from cell cultures or laboratory animals for genomic extraction to amplify target genes. Synthetic genes have gained a key role in recombinant protein production. For the first time, we demonstrated the production of the recombinant protein of the T. gondii dense granular antigen 8 (TgGRA8) gene based on commercial gene synthesis. Recombinant TgGRA8 plasmids were successfully expressed in an Escherichia coli system. The recombinant protein was affinity-purified and characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Furthermore, the diagnostic potential of the recombinant protein was assessed using 306 field serum samples from goats via indirect enzyme-linked immunosorbent assay (iELISA) and the latex agglutination test (LAT). Results Western blotting using known positive serum samples from goats identified a single antigen at the expected molecular weight of TgGRA8 (27 kDa). iELISA illustrated that 15.40% of goat samples were positive for T. gondii-specific IgG antibodies. In addition, TgGRA8 provided high sensitivity and specificity, with significant concordance (91.83) and kappa values (0.69) compared with the results obtained using LAT. Conclusion Our findings highlight the production of a recombinant protein from a synthetic TgGRA8 gene and the ability to detect T. gondii infection in field samples. The sensitivity and specificity of TgGRA8 demonstrated that this protein could be a good serological marker for detecting specific IgG in goat sera.

Can Osteopontin Be Considered a Biomarker for Endometriosis?

2017 ◽  
Vol 68 (9) ◽  
pp. 2132-2134
Author(s):  
Daniela Roxana Albu (Matasariu) ◽  
Elena Mihalceanu ◽  
Alina Pangal ◽  
Carmen Vulpoi ◽  
Mircea Onofriescu ◽  
...  
Keyword(s):  
Serum Level ◽  
Immunosorbent Assay ◽  
Pelvic Pain ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Multifactorial Disease ◽  
Elisa Kit ◽  
Healthy Women ◽  
Progesterone Treatment ◽  
Large Dispersion

Endometriosis is a multifactorial disease that is manifested by infertility and pelvic pain. The purpose of the study was to evaluate the effect of progesterone treatment on the serum level of osteopontin, a multipotent cytokine, in patients with endometriosis. The study was prospective and we evaluated osteopontin levels that were measured in the serum of 40 patients with endometriosis and 12 healthy women using a standardized Enzyme-Linked Immunosorbent Assay (ELISA) kit. Osteopontin seric levels were lower in endometriosis patients and increased after progesterone treatment. Because of the large dispersion of data even in the control group, we find the association between osteopontin and endometriosis questionable.

scholarly journals Presence of anti-Toxoplasma antibodies in humans and their cats in the urban zone of Guadalajara

1999 ◽  
Vol 32 (5) ◽  
pp. 483-488 ◽  
Author(s):  
María de la Luz Galván Ramírez ◽  
Guillermo Sánchez Vargas ◽  
Marcos Vielma Sandoval ◽  
Juan Luis Soto Mancilla
Keyword(s):  
Risk Factors ◽  
Toxoplasma Gondii ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Toxoplasma Infection ◽  
Urban Zone ◽  
Lack Of Control ◽  
Iga Antibodies

Cats are the definitive hosts of Toxoplasma gondii. Infected cats excrete oocysts in their feces, infecting humans and other animals. The objective of the present study was to determine the presence of anti-Toxoplasma antibodies in cat owners and their pets, and determine if there was a relationship between Toxoplasma infection and humans who live with infected cats. IgG anti-Toxoplasma antibodies in sera of 59 cat owners were determined by enzyme-linked immunosorbent assay (ELISA), in 24 sera from their cats, IgG, IgM, and IgA antibodies were found using Burney's ELISA. Thirty-eight (64%) of 59 cat owners were positive to IgG anti-Toxoplasma. Seropositivity for cats was 70.8% IgG, 8.3% IgM, and 62.5% IgA. Cohabitation with cats infected by T. gondii, feeding with leftovers or raw viscera, and lack of control over how their feces were handled are risk factors conducive for humans to become infected by T. gondii.

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