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Diagnostic value of recombinant nanoluciferase fused Toxoplasma gondii antigens in Luciferase-linked Antibody Capture Assay (LACA) for Toxoplasma infection in pigs
Toxoplasmosis is a widespread protozoan zoonosis. Since ingesting undercooked meat harboring Toxoplasma gondii cyst is considered one of the major transmission routes to humans, the screening of T. gondii in meat-producing animals can reduce the risk of food-borne toxoplasmosis in humans. Among sero...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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The Japanese Society of Veterinary Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9353080/ https://www.ncbi.nlm.nih.gov/pubmed/35584943 http://dx.doi.org/10.1292/jvms.22-0099 |
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author | DUONG, Hieu Duc TANIGUCHI, Yuji TAKASHIMA, Yasuhiro SEKIGUCHI, Satoshi AYE, Khin Myo AHMADI, Parnian BUI, Linh Khanh IRIE, Takao NAGAYASU, Eiji YOSHIDA, Ayako |
author_facet | DUONG, Hieu Duc TANIGUCHI, Yuji TAKASHIMA, Yasuhiro SEKIGUCHI, Satoshi AYE, Khin Myo AHMADI, Parnian BUI, Linh Khanh IRIE, Takao NAGAYASU, Eiji YOSHIDA, Ayako |
author_sort | DUONG, Hieu Duc |
collection | PubMed |
description | Toxoplasmosis is a widespread protozoan zoonosis. Since ingesting undercooked meat harboring Toxoplasma gondii cyst is considered one of the major transmission routes to humans, the screening of T. gondii in meat-producing animals can reduce the risk of food-borne toxoplasmosis in humans. Among serological diagnostic methods, Luciferase-linked Antibody Capture Assay (LACA) has been found to be a promising platform with high sensitivity and specificity. In this study, we aimed to evaluate recombinant nanoluciferase fused-T. gondii antigens (rNluc-GRA6, rNluc-GRA7, rNluc-GRA8 and rNluc-BAG1) for their potential use in LACA for pigs. As a result, the sensitivity of GRA6-, GRA7-, GRA8- and BAG1-LACA were 70.0%, 80.0%, 80.0% and 30.0% with specificity 87.0%, 81.5%, 74.1% and 50.0%, respectively. The cocktail LACA using a mixture of rNluc-GRA6, rNluc-GRA7 and rNluc-GRA8 indicated higher sensitivity (90.0%) and a similar specificity (96.3%) in comparison with the commercial ELISA kit. Compared to the Dye-Test as a reference test, cocktail LACA showed strong agreement (kappa value=0.811) when we assessed pig sera collected at the slaughterhouse. In addition, we also successfully established the rapid LACA format for the detection of Toxoplasma infection in pigs (called Rapid-LACA) in which the test could be performed within 30 min. In Rapid-LACA, the protein A pre-coated/blocked plates could be preserved at −30°C, 4°C or room temperature conditions for at least two months without compromising on the quality of assay. |
format | Online Article Text |
id | pubmed-9353080 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Japanese Society of Veterinary Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-93530802022-08-06 Diagnostic value of recombinant nanoluciferase fused Toxoplasma gondii antigens in Luciferase-linked Antibody Capture Assay (LACA) for Toxoplasma infection in pigs DUONG, Hieu Duc TANIGUCHI, Yuji TAKASHIMA, Yasuhiro SEKIGUCHI, Satoshi AYE, Khin Myo AHMADI, Parnian BUI, Linh Khanh IRIE, Takao NAGAYASU, Eiji YOSHIDA, Ayako J Vet Med Sci Parasitology Toxoplasmosis is a widespread protozoan zoonosis. Since ingesting undercooked meat harboring Toxoplasma gondii cyst is considered one of the major transmission routes to humans, the screening of T. gondii in meat-producing animals can reduce the risk of food-borne toxoplasmosis in humans. Among serological diagnostic methods, Luciferase-linked Antibody Capture Assay (LACA) has been found to be a promising platform with high sensitivity and specificity. In this study, we aimed to evaluate recombinant nanoluciferase fused-T. gondii antigens (rNluc-GRA6, rNluc-GRA7, rNluc-GRA8 and rNluc-BAG1) for their potential use in LACA for pigs. As a result, the sensitivity of GRA6-, GRA7-, GRA8- and BAG1-LACA were 70.0%, 80.0%, 80.0% and 30.0% with specificity 87.0%, 81.5%, 74.1% and 50.0%, respectively. The cocktail LACA using a mixture of rNluc-GRA6, rNluc-GRA7 and rNluc-GRA8 indicated higher sensitivity (90.0%) and a similar specificity (96.3%) in comparison with the commercial ELISA kit. Compared to the Dye-Test as a reference test, cocktail LACA showed strong agreement (kappa value=0.811) when we assessed pig sera collected at the slaughterhouse. In addition, we also successfully established the rapid LACA format for the detection of Toxoplasma infection in pigs (called Rapid-LACA) in which the test could be performed within 30 min. In Rapid-LACA, the protein A pre-coated/blocked plates could be preserved at −30°C, 4°C or room temperature conditions for at least two months without compromising on the quality of assay. The Japanese Society of Veterinary Science 2022-05-18 2022-07 /pmc/articles/PMC9353080/ /pubmed/35584943 http://dx.doi.org/10.1292/jvms.22-0099 Text en ©2022 The Japanese Society of Veterinary Science https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/) |
spellingShingle | Parasitology DUONG, Hieu Duc TANIGUCHI, Yuji TAKASHIMA, Yasuhiro SEKIGUCHI, Satoshi AYE, Khin Myo AHMADI, Parnian BUI, Linh Khanh IRIE, Takao NAGAYASU, Eiji YOSHIDA, Ayako Diagnostic value of recombinant nanoluciferase fused Toxoplasma gondii antigens in Luciferase-linked Antibody Capture Assay (LACA) for Toxoplasma infection in pigs |
title | Diagnostic value of recombinant nanoluciferase fused Toxoplasma gondii antigens in Luciferase-linked Antibody Capture Assay (LACA) for
Toxoplasma infection in pigs |
title_full | Diagnostic value of recombinant nanoluciferase fused Toxoplasma gondii antigens in Luciferase-linked Antibody Capture Assay (LACA) for
Toxoplasma infection in pigs |
title_fullStr | Diagnostic value of recombinant nanoluciferase fused Toxoplasma gondii antigens in Luciferase-linked Antibody Capture Assay (LACA) for
Toxoplasma infection in pigs |
title_full_unstemmed | Diagnostic value of recombinant nanoluciferase fused Toxoplasma gondii antigens in Luciferase-linked Antibody Capture Assay (LACA) for
Toxoplasma infection in pigs |
title_short | Diagnostic value of recombinant nanoluciferase fused Toxoplasma gondii antigens in Luciferase-linked Antibody Capture Assay (LACA) for
Toxoplasma infection in pigs |
title_sort | diagnostic value of recombinant nanoluciferase fused toxoplasma gondii antigens in luciferase-linked antibody capture assay (laca) for
toxoplasma infection in pigs |
topic | Parasitology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9353080/ https://www.ncbi.nlm.nih.gov/pubmed/35584943 http://dx.doi.org/10.1292/jvms.22-0099 |
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