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Article

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

1
Laboratory of Microbiology, Division of Disease Control, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan
2
Global Station for Zoonosis Control, Global Institute for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo 001-0020, Japan
3
Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan
4
Department of Molecular Biology, Princeton University, Washington Road, Princeton, NJ 08540, USA
5
Institute of Virology and Cell Biology, University of Lübeck, D-23562 Lübeck, Germany
*
Author to whom correspondence should be addressed.
Pathogens 2020, 9(3), 188; https://doi.org/10.3390/pathogens9030188
Received: 28 January 2020 / Revised: 2 March 2020 / Accepted: 3 March 2020 / Published: 4 March 2020
(This article belongs to the Special Issue Classical Swine Fever)
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. View Full-Text
Keywords: border disease; bovine viral diarrhea; classical swine fever; pestivirus; serum neutralization test; reporter virus border disease; bovine viral diarrhea; classical swine fever; pestivirus; serum neutralization test; reporter virus
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MDPI and ACS Style

Tetsuo, M.; Matsuno, K.; Tamura, T.; Fukuhara, T.; Kim, T.; Okamatsu, M.; Tautz, N.; Matsuura, Y.; Sakoda, Y. Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag. Pathogens 2020, 9, 188. https://doi.org/10.3390/pathogens9030188

AMA Style

Tetsuo M, Matsuno K, Tamura T, Fukuhara T, Kim T, Okamatsu M, Tautz N, Matsuura Y, Sakoda Y. Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag. Pathogens. 2020; 9(3):188. https://doi.org/10.3390/pathogens9030188

Chicago/Turabian Style

Tetsuo, Madoka, Keita Matsuno, Tomokazu Tamura, Takasuke Fukuhara, Taksoo Kim, Masatoshi Okamatsu, Norbert Tautz, Yoshiharu Matsuura, and Yoshihiro Sakoda. 2020. "Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag" Pathogens 9, no. 3: 188. https://doi.org/10.3390/pathogens9030188

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