Development of a high-throughput serum neutralization test using recombinant pestiviruses possessing a small reporter tag

Madoka Tetsuo; Keita Matsuno; Tomokazu Tamura; Takasuke Fukuhara; Taksoo Kim; Masatoshi Okamatsu; Norbert Tautz; Yoshiharu Matsuura; Yoshihiro Sakoda

doi:10.3390/pathogens9030188

Development of a high-throughput serum neutralization test using recombinant pestiviruses possessing a small reporter tag

Madoka Tetsuo, Keita Matsuno, Tomokazu Tamura, Takasuke Fukuhara, Taksoo Kim, Masatoshi Okamatsu, Norbert Tautz, Yoshiharu Matsuura, Yoshihiro Sakoda^*

^*Corresponding author for this work

Institute of Virology and Cell Biology

18 Citations (Scopus)

Abstract

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.

Original language	English
Article number	188
Journal	Pathogens
Volume	9
Issue number	3
DOIs	https://doi.org/10.3390/pathogens9030188
Publication status	Published - 03.2020

Funding

Funding: This study was partially funded by a Grant-in-Aid for Scientific Research on Innovative Areas from the Ministry of Education, Culture, Science, Sports, and Technology (MEXT) of Japan (MEXT KAKENHI Grant Numbers: JP19H03115, JP16H06429, JP16K21723, and JP16H06431); and the Japan Initiative for Global Research Network on Infectious Diseases (J-GRID) (Grant Number: PJ18fm0108008) from the Japan Agency for Medical Research and Development (AMED).

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Research Areas and Centers

Academic Focus: Center for Infection and Inflammation Research (ZIEL)

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Cite this

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title = "Development of a high-throughput serum neutralization test using recombinant pestiviruses possessing a small reporter tag",

abstract = "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.",

author = "Madoka Tetsuo and Keita Matsuno and Tomokazu Tamura and Takasuke Fukuhara and Taksoo Kim and Masatoshi Okamatsu and Norbert Tautz and Yoshiharu Matsuura and Yoshihiro Sakoda",

note = "Funding Information: Funding: This study was partially funded by a Grant-in-Aid for Scientific Research on Innovative Areas from the Ministry of Education, Culture, Science, Sports, and Technology (MEXT) of Japan (MEXT KAKENHI Grant Numbers: JP19H03115, JP16H06429, JP16K21723, and JP16H06431); and the Japan Initiative for Global Research Network on Infectious Diseases (J-GRID) (Grant Number: PJ18fm0108008) from the Japan Agency for Medical Research and Development (AMED). Publisher Copyright: {\textcopyright} 2020 by the authors. Licensee MDPI, Basel, Switzerland. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

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AU - Tetsuo, Madoka

AU - Matsuno, Keita

AU - Tamura, Tomokazu

AU - Fukuhara, Takasuke

AU - Kim, Taksoo

AU - Okamatsu, Masatoshi

AU - Tautz, Norbert

AU - Matsuura, Yoshiharu

AU - Sakoda, Yoshihiro

N1 - Funding Information: Funding: This study was partially funded by a Grant-in-Aid for Scientific Research on Innovative Areas from the Ministry of Education, Culture, Science, Sports, and Technology (MEXT) of Japan (MEXT KAKENHI Grant Numbers: JP19H03115, JP16H06429, JP16K21723, and JP16H06431); and the Japan Initiative for Global Research Network on Infectious Diseases (J-GRID) (Grant Number: PJ18fm0108008) from the Japan Agency for Medical Research and Development (AMED). Publisher Copyright: © 2020 by the authors. Licensee MDPI, Basel, Switzerland. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.

PY - 2020/3

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N2 - 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.

AB - 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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ER -

Development of a high-throughput serum neutralization test using recombinant pestiviruses possessing a small reporter tag

Abstract

Funding

UN SDGs

Research Areas and Centers

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