TY - JOUR
T1 - Development of a high-throughput serum neutralization test using recombinant pestiviruses possessing a small reporter tag
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
Y1 - 2020/3
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.
UR - http://www.scopus.com/inward/record.url?scp=85082405891&partnerID=8YFLogxK
U2 - 10.3390/pathogens9030188
DO - 10.3390/pathogens9030188
M3 - Journal articles
AN - SCOPUS:85082405891
VL - 9
JO - Pathogens
JF - Pathogens
IS - 3
M1 - 188
ER -
