Development of an indirect ELISA for detecting Toxoplasma gondii IgG antibodies based on a recombinant TgIMP1 protein

Toxoplasma gondii ( T . gondii ) is widely spread around the world, which can cause serious harm to immunosuppressed patients. Currently, the commercial test kits are poor at assessing T . gondii infection and vaccine effectiveness, making an urgent need to exploit effective enzyme-linked immunosorb...

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Published inPLoS neglected tropical diseases Vol. 18; no. 8; p. e0012421
Main Authors Dong, Hongjie, Zhang, Junmei, Wang, Qi, Shen, Yanmei, Zhou, Beibei, Dai, Lisha, Zhu, Wenju, Sun, Hang, Xie, Xiaoman, Xie, Huanhuan, Xu, Chao, Zhao, Guihua, Yin, Kun
Format Journal Article
LanguageEnglish
Published United States Public Library of Science 14.08.2024
Public Library of Science (PLoS)
Subjects
Analysis
Animals
Antibodies, Protozoan - blood
Antigens, Protozoan - genetics
Antigens, Protozoan - immunology
Biology and Life Sciences
Care and treatment
Diagnosis
Dosage and administration
Engineering and Technology
Enzyme-linked immunosorbent assay
Enzyme-Linked Immunosorbent Assay - methods
Escherichia coli - genetics
Humans
Immunoglobulin G
Immunoglobulin G - blood
Medicine and Health Sciences
Physical Sciences
Protozoan Proteins - genetics
Protozoan Proteins - immunology
Recombinant proteins
Recombinant Proteins - genetics
Recombinant Proteins - immunology
Research and Analysis Methods
Risk factors
Sensitivity and Specificity
Toxoplasma - genetics
Toxoplasma - immunology
Toxoplasmosis
Toxoplasmosis - diagnosis
Toxoplasmosis - immunology
Toxoplasmosis - parasitology
China
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Abstract Toxoplasma gondii ( T . gondii ) is widely spread around the world, which can cause serious harm to immunosuppressed patients. Currently, the commercial test kits are poor at assessing T . gondii infection and vaccine effectiveness, making an urgent need to exploit effective enzyme-linked immunosorbent assay with great performance to compensate for this deficiency. Here, the TgIMP1 recombinant protein was expressed in E . coli BL(21) cells. The TgIMP1 was purified with affinity chromatography and the reactivity was retained with anti-TgIMP1 antibodies. The TgIMP1 was then used to develop an indirect ELISA (IMP1-iELISA) and the reaction conditions of IMP1-iELISA were optimized. As a result, the cut-off value was determined to be 0.2833 by analyzing the OD 450nm values of forty T . gondii -negative sera. The coefficient of variation of 6 T . gondii -positive sera within and between runs were both less than 10%. The IMP1-iELISA was non-cross-reactive with the sera of cytomegalovirus, herpes virus, rubella virus, Cryptosporidium spp., Theileria spp., Neospora spp. and Plasmodium spp.. Furthermore, the sensitivity and specificity of IMP1-iELISA were 98.9% and 96.7%, respectively, based on testing 150 serum samples. The results suggest that this IMP1-iELISA is specific, sensitive, repeatable and can be applied to the detection of T. gondii infections in the medical and health industries.
AbstractList Toxoplasma gondii (T. gondii) is widely spread around the world, which can cause serious harm to immunosuppressed patients. Currently, the commercial test kits are poor at assessing T. gondii infection and vaccine effectiveness, making an urgent need to exploit effective enzyme-linked immunosorbent assay with great performance to compensate for this deficiency. Here, the TgIMP1 recombinant protein was expressed in E. coli BL(21) cells. The TgIMP1 was purified with affinity chromatography and the reactivity was retained with anti-TgIMP1 antibodies. The TgIMP1 was then used to develop an indirect ELISA (IMP1-iELISA) and the reaction conditions of IMP1-iELISA were optimized. As a result, the cut-off value was determined to be 0.2833 by analyzing the OD450nm values of forty T. gondii-negative sera. The coefficient of variation of 6 T. gondii-positive sera within and between runs were both less than 10%. The IMP1-iELISA was non-cross-reactive with the sera of cytomegalovirus, herpes virus, rubella virus, Cryptosporidium spp., Theileria spp., Neospora spp. and Plasmodium spp.. Furthermore, the sensitivity and specificity of IMP1-iELISA were 98.9% and 96.7%, respectively, based on testing 150 serum samples. The results suggest that this IMP1-iELISA is specific, sensitive, repeatable and can be applied to the detection of T. gondii infections in the medical and health industries.
Toxoplasma gondii ( T . gondii ) is widely spread around the world, which can cause serious harm to immunosuppressed patients. Currently, the commercial test kits are poor at assessing T . gondii infection and vaccine effectiveness, making an urgent need to exploit effective enzyme-linked immunosorbent assay with great performance to compensate for this deficiency. Here, the TgIMP1 recombinant protein was expressed in E . coli BL(21) cells. The TgIMP1 was purified with affinity chromatography and the reactivity was retained with anti-TgIMP1 antibodies. The TgIMP1 was then used to develop an indirect ELISA (IMP1-iELISA) and the reaction conditions of IMP1-iELISA were optimized. As a result, the cut-off value was determined to be 0.2833 by analyzing the OD 450nm values of forty T . gondii -negative sera. The coefficient of variation of 6 T . gondii -positive sera within and between runs were both less than 10%. The IMP1-iELISA was non-cross-reactive with the sera of cytomegalovirus, herpes virus, rubella virus, Cryptosporidium spp., Theileria spp., Neospora spp. and Plasmodium spp.. Furthermore, the sensitivity and specificity of IMP1-iELISA were 98.9% and 96.7%, respectively, based on testing 150 serum samples. The results suggest that this IMP1-iELISA is specific, sensitive, repeatable and can be applied to the detection of T. gondii infections in the medical and health industries. Toxoplasma gondii is an extremely widespread and a worldwide health problem protozoon, which can induce acute and chronic infection and cause severe toxoplasmosis. However, there is still no effective vaccine to prevent infection, drugs used to treat the acute infected individuals have a lot of limitations, and no drugs to treat chronic infected individuals. Indirect ELISA, as a commonly used clinical test with satisfactory detection performance and easy implementation for high throughput testing, has been used to detect various pathogenic microorganisms. Immune mapped protein1 (IMP1) has been identified as a protein that induces a strong immune response in the host. In this study, we obtained IMP1 protein with high purity and used it as coated antigen to prepare an indirect ELISA reagent, and then verified its detection performance (sensitivity, specificity, repeatability). This study enriches the diagnosis of toxoplasmosis and provides strong technical support for epidemiological investigation and transmission control of toxoplasmosis.
Toxoplasma gondii ( T . gondii ) is widely spread around the world, which can cause serious harm to immunosuppressed patients. Currently, the commercial test kits are poor at assessing T . gondii infection and vaccine effectiveness, making an urgent need to exploit effective enzyme-linked immunosorbent assay with great performance to compensate for this deficiency. Here, the TgIMP1 recombinant protein was expressed in E . coli BL(21) cells. The TgIMP1 was purified with affinity chromatography and the reactivity was retained with anti-TgIMP1 antibodies. The TgIMP1 was then used to develop an indirect ELISA (IMP1-iELISA) and the reaction conditions of IMP1-iELISA were optimized. As a result, the cut-off value was determined to be 0.2833 by analyzing the OD 450nm values of forty T . gondii -negative sera. The coefficient of variation of 6 T . gondii -positive sera within and between runs were both less than 10%. The IMP1-iELISA was non-cross-reactive with the sera of cytomegalovirus, herpes virus, rubella virus, Cryptosporidium spp., Theileria spp., Neospora spp. and Plasmodium spp.. Furthermore, the sensitivity and specificity of IMP1-iELISA were 98.9% and 96.7%, respectively, based on testing 150 serum samples. The results suggest that this IMP1-iELISA is specific, sensitive, repeatable and can be applied to the detection of T. gondii infections in the medical and health industries.
Toxoplasma gondii (T. gondii) is widely spread around the world, which can cause serious harm to immunosuppressed patients. Currently, the commercial test kits are poor at assessing T. gondii infection and vaccine effectiveness, making an urgent need to exploit effective enzyme-linked immunosorbent assay with great performance to compensate for this deficiency. Here, the TgIMP1 recombinant protein was expressed in E. coli BL(21) cells. The TgIMP1 was purified with affinity chromatography and the reactivity was retained with anti-TgIMP1 antibodies. The TgIMP1 was then used to develop an indirect ELISA (IMP1-iELISA) and the reaction conditions of IMP1-iELISA were optimized. As a result, the cut-off value was determined to be 0.2833 by analyzing the OD.sub.450nm values of forty T. gondii-negative sera. The coefficient of variation of 6 T. gondii-positive sera within and between runs were both less than 10%. The IMP1-iELISA was non-cross-reactive with the sera of cytomegalovirus, herpes virus, rubella virus, Cryptosporidium spp., Theileria spp., Neospora spp. and Plasmodium spp.. Furthermore, the sensitivity and specificity of IMP1-iELISA were 98.9% and 96.7%, respectively, based on testing 150 serum samples. The results suggest that this IMP1-iELISA is specific, sensitive, repeatable and can be applied to the detection of T. gondii infections in the medical and health industries.
Toxoplasma gondii (T. gondii) is widely spread around the world, which can cause serious harm to immunosuppressed patients. Currently, the commercial test kits are poor at assessing T. gondii infection and vaccine effectiveness, making an urgent need to exploit effective enzyme-linked immunosorbent assay with great performance to compensate for this deficiency. Here, the TgIMP1 recombinant protein was expressed in E. coli BL(21) cells. The TgIMP1 was purified with affinity chromatography and the reactivity was retained with anti-TgIMP1 antibodies. The TgIMP1 was then used to develop an indirect ELISA (IMP1-iELISA) and the reaction conditions of IMP1-iELISA were optimized. As a result, the cut-off value was determined to be 0.2833 by analyzing the OD450nm values of forty T. gondii-negative sera. The coefficient of variation of 6 T. gondii-positive sera within and between runs were both less than 10%. The IMP1-iELISA was non-cross-reactive with the sera of cytomegalovirus, herpes virus, rubella virus, Cryptosporidium spp., Theileria spp., Neospora spp. and Plasmodium spp.. Furthermore, the sensitivity and specificity of IMP1-iELISA were 98.9% and 96.7%, respectively, based on testing 150 serum samples. The results suggest that this IMP1-iELISA is specific, sensitive, repeatable and can be applied to the detection of T. gondii infections in the medical and health industries.Toxoplasma gondii (T. gondii) is widely spread around the world, which can cause serious harm to immunosuppressed patients. Currently, the commercial test kits are poor at assessing T. gondii infection and vaccine effectiveness, making an urgent need to exploit effective enzyme-linked immunosorbent assay with great performance to compensate for this deficiency. Here, the TgIMP1 recombinant protein was expressed in E. coli BL(21) cells. The TgIMP1 was purified with affinity chromatography and the reactivity was retained with anti-TgIMP1 antibodies. The TgIMP1 was then used to develop an indirect ELISA (IMP1-iELISA) and the reaction conditions of IMP1-iELISA were optimized. As a result, the cut-off value was determined to be 0.2833 by analyzing the OD450nm values of forty T. gondii-negative sera. The coefficient of variation of 6 T. gondii-positive sera within and between runs were both less than 10%. The IMP1-iELISA was non-cross-reactive with the sera of cytomegalovirus, herpes virus, rubella virus, Cryptosporidium spp., Theileria spp., Neospora spp. and Plasmodium spp.. Furthermore, the sensitivity and specificity of IMP1-iELISA were 98.9% and 96.7%, respectively, based on testing 150 serum samples. The results suggest that this IMP1-iELISA is specific, sensitive, repeatable and can be applied to the detection of T. gondii infections in the medical and health industries.
Audience Academic
Author Zhao, Guihua
Zhang, Junmei
Xie, Xiaoman
Sun, Hang
Yin, Kun
Dai, Lisha
Wang, Qi
Dong, Hongjie
Shen, Yanmei
Xu, Chao
Xie, Huanhuan
Zhou, Beibei
Zhu, Wenju
AuthorAffiliation 2 Digestive Disease Hospital of Shandong First Medical University, Shandong First Medical University & Shandong Academy of Medical Sciences, Jining, People’s Republic of China
University of Texas at El Paso, UNITED STATES OF AMERICA
1 Shandong Institute of Parasitic Diseases, Shandong First Medical University & Shandong Academy of Medical Sciences, Jining, People’s Republic of China
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– name: University of Texas at El Paso, UNITED STATES OF AMERICA
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Snippet Toxoplasma gondii ( T . gondii ) is widely spread around the world, which can cause serious harm to immunosuppressed patients. Currently, the commercial test...
Toxoplasma gondii (T. gondii) is widely spread around the world, which can cause serious harm to immunosuppressed patients. Currently, the commercial test kits...
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SubjectTerms Analysis
Animals
Antibodies, Protozoan - blood
Antigens, Protozoan - genetics
Antigens, Protozoan - immunology
Biology and Life Sciences
Care and treatment
Diagnosis
Dosage and administration
Engineering and Technology
Enzyme-linked immunosorbent assay
Enzyme-Linked Immunosorbent Assay - methods
Escherichia coli - genetics
Humans
Immunoglobulin G
Immunoglobulin G - blood
Medicine and Health Sciences
Physical Sciences
Protozoan Proteins - genetics
Protozoan Proteins - immunology
Recombinant proteins
Recombinant Proteins - genetics
Recombinant Proteins - immunology
Research and Analysis Methods
Risk factors
Sensitivity and Specificity
Toxoplasma - genetics
Toxoplasma - immunology
Toxoplasmosis
Toxoplasmosis - diagnosis
Toxoplasmosis - immunology
Toxoplasmosis - parasitology
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Title Development of an indirect ELISA for detecting Toxoplasma gondii IgG antibodies based on a recombinant TgIMP1 protein
URI https://www.ncbi.nlm.nih.gov/pubmed/39141677
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