Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: DUONG, Hieu Duc, TANIGUCHI, Yuji, TAKASHIMA, Yasuhiro, SEKIGUCHI, Satoshi, AYE, Khin Myo, AHMADI, Parnian, BUI, Linh Khanh, IRIE, Takao, NAGAYASU, Eiji, YOSHIDA, Ayako
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Japanese Society of Veterinary Science 2022
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
_version_ 1784762795802230784
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
work_keys_str_mv AT duonghieuduc diagnosticvalueofrecombinantnanoluciferasefusedtoxoplasmagondiiantigensinluciferaselinkedantibodycaptureassaylacafortoxoplasmainfectioninpigs
AT taniguchiyuji diagnosticvalueofrecombinantnanoluciferasefusedtoxoplasmagondiiantigensinluciferaselinkedantibodycaptureassaylacafortoxoplasmainfectioninpigs
AT takashimayasuhiro diagnosticvalueofrecombinantnanoluciferasefusedtoxoplasmagondiiantigensinluciferaselinkedantibodycaptureassaylacafortoxoplasmainfectioninpigs
AT sekiguchisatoshi diagnosticvalueofrecombinantnanoluciferasefusedtoxoplasmagondiiantigensinluciferaselinkedantibodycaptureassaylacafortoxoplasmainfectioninpigs
AT ayekhinmyo diagnosticvalueofrecombinantnanoluciferasefusedtoxoplasmagondiiantigensinluciferaselinkedantibodycaptureassaylacafortoxoplasmainfectioninpigs
AT ahmadiparnian diagnosticvalueofrecombinantnanoluciferasefusedtoxoplasmagondiiantigensinluciferaselinkedantibodycaptureassaylacafortoxoplasmainfectioninpigs
AT builinhkhanh diagnosticvalueofrecombinantnanoluciferasefusedtoxoplasmagondiiantigensinluciferaselinkedantibodycaptureassaylacafortoxoplasmainfectioninpigs
AT irietakao diagnosticvalueofrecombinantnanoluciferasefusedtoxoplasmagondiiantigensinluciferaselinkedantibodycaptureassaylacafortoxoplasmainfectioninpigs
AT nagayasueiji diagnosticvalueofrecombinantnanoluciferasefusedtoxoplasmagondiiantigensinluciferaselinkedantibodycaptureassaylacafortoxoplasmainfectioninpigs
AT yoshidaayako diagnosticvalueofrecombinantnanoluciferasefusedtoxoplasmagondiiantigensinluciferaselinkedantibodycaptureassaylacafortoxoplasmainfectioninpigs