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Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag

A serum neutralization test (SNT) is an essential method for the serological diagnosis of pestivirus infections, including classical swine fever, because of the cross reactivity of antibodies against pestiviruses and the non-quantitative properties of antibodies in an enzyme-linked immunosorbent ass...

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Autores principales: Tetsuo, Madoka, Matsuno, Keita, Tamura, Tomokazu, Fukuhara, Takasuke, Kim, Taksoo, Okamatsu, Masatoshi, Tautz, Norbert, Matsuura, Yoshiharu, Sakoda, Yoshihiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7157198/
https://www.ncbi.nlm.nih.gov/pubmed/32143534
http://dx.doi.org/10.3390/pathogens9030188
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author Tetsuo, Madoka
Matsuno, Keita
Tamura, Tomokazu
Fukuhara, Takasuke
Kim, Taksoo
Okamatsu, Masatoshi
Tautz, Norbert
Matsuura, Yoshiharu
Sakoda, Yoshihiro
author_facet Tetsuo, Madoka
Matsuno, Keita
Tamura, Tomokazu
Fukuhara, Takasuke
Kim, Taksoo
Okamatsu, Masatoshi
Tautz, Norbert
Matsuura, Yoshiharu
Sakoda, Yoshihiro
author_sort Tetsuo, Madoka
collection PubMed
description A serum neutralization test (SNT) is an essential method for the serological diagnosis of pestivirus infections, including classical swine fever, because of the cross reactivity of antibodies against pestiviruses and the non-quantitative properties of antibodies in an enzyme-linked immunosorbent assay. In conventional SNTs, an immunoperoxidase assay or observation of cytopathic effect after incubation for 3 to 7 days is needed to determine the SNT titer, which requires labor-intensive or time-consuming procedures. Therefore, a new SNT, based on the luciferase system and using classical swine fever virus, bovine viral diarrhea virus, and border disease virus possessing the 11-amino-acid subunit derived from NanoLuc luciferase was developed and evaluated; this approach enabled the rapid and easy determination of the SNT titer using a luminometer. In the new method, SNT titers can be determined tentatively at 2 days post-infection (dpi) and are comparable to those obtained by conventional SNTs at 3 or 4 dpi. In conclusion, the luciferase-based SNT can replace conventional SNTs as a high-throughput antibody test for pestivirus infections.
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spelling pubmed-71571982020-05-01 Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag Tetsuo, Madoka Matsuno, Keita Tamura, Tomokazu Fukuhara, Takasuke Kim, Taksoo Okamatsu, Masatoshi Tautz, Norbert Matsuura, Yoshiharu Sakoda, Yoshihiro Pathogens Article A serum neutralization test (SNT) is an essential method for the serological diagnosis of pestivirus infections, including classical swine fever, because of the cross reactivity of antibodies against pestiviruses and the non-quantitative properties of antibodies in an enzyme-linked immunosorbent assay. In conventional SNTs, an immunoperoxidase assay or observation of cytopathic effect after incubation for 3 to 7 days is needed to determine the SNT titer, which requires labor-intensive or time-consuming procedures. Therefore, a new SNT, based on the luciferase system and using classical swine fever virus, bovine viral diarrhea virus, and border disease virus possessing the 11-amino-acid subunit derived from NanoLuc luciferase was developed and evaluated; this approach enabled the rapid and easy determination of the SNT titer using a luminometer. In the new method, SNT titers can be determined tentatively at 2 days post-infection (dpi) and are comparable to those obtained by conventional SNTs at 3 or 4 dpi. In conclusion, the luciferase-based SNT can replace conventional SNTs as a high-throughput antibody test for pestivirus infections. MDPI 2020-03-04 /pmc/articles/PMC7157198/ /pubmed/32143534 http://dx.doi.org/10.3390/pathogens9030188 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Tetsuo, Madoka
Matsuno, Keita
Tamura, Tomokazu
Fukuhara, Takasuke
Kim, Taksoo
Okamatsu, Masatoshi
Tautz, Norbert
Matsuura, Yoshiharu
Sakoda, Yoshihiro
Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag
title Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag
title_full Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag
title_fullStr Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag
title_full_unstemmed Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag
title_short Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag
title_sort development of a high-throughput serum neutralization test using recombinant pestiviruses possessing a small reporter tag
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7157198/
https://www.ncbi.nlm.nih.gov/pubmed/32143534
http://dx.doi.org/10.3390/pathogens9030188
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