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

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Title: Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag
Authors: Tetsuo, Madoka Browse this author
Matsuno, Keita Browse this author →KAKEN DB
Tamura, Tomokazu Browse this author
Fukuhara, Takasuke Browse this author →KAKEN DB
Kim, Taksoo Browse this author
Okamatsu, Masatoshi Browse this author →KAKEN DB
Tautz, Norbert Browse this author
Matsuura, Yoshiharu Browse this author →KAKEN DB
Sakoda, Yoshihiro Browse this author →KAKEN DB
Keywords: border disease
bovine viral diarrhea
classical swine fever
serum neutralization test
reporter virus
Issue Date: Mar-2020
Publisher: MDPI
Journal Title: Pathogens
Volume: 9
Issue: 3
Start Page: 188
Publisher DOI: 10.3390/pathogens9030188
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.
Type: article
Appears in Collections:国際連携研究教育局 : GI-CoRE (Global Institution for Collaborative Research and Education : GI-CoRE) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
獣医学院・獣医学研究院 (Graduate School of Veterinary Medicine / Faculty of Veterinary Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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