Establishment of the Foot-and-Mouth Disease Virus Type Asia1 Expressing the HiBiT Protein: A Useful Tool for a NanoBiT Split Luciferase Assay
Giyoun Cho,
Hyejin Kim,
Dong-Wan Kim,
Seong Yun Hwang,
Ji-Hyeon Hwang,
Yeon Rae Chae,
Yoon-Hee Lee,
Ok-Mi Jeong,
Jung-Won Park,
Sung-Han Park,
Jong-Hyeon Park
Affiliations
Giyoun Cho
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Hyejin Kim
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Dong-Wan Kim
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Seong Yun Hwang
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Ji-Hyeon Hwang
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Yeon Rae Chae
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Yoon-Hee Lee
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Ok-Mi Jeong
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Jung-Won Park
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Sung-Han Park
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Jong-Hyeon Park
Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177, Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea
Foot-and-mouth disease virus (FMDV) is a highly contagious virus that affects cloven-hoofed animals and causes severe economic losses in the livestock industry. Given that this high-risk pathogen has to be handled in a biosafety level (BSL)-3 facility for safety reasons and the limited availability of BSL-3 laboratories, experiments on FMDV call for more attention. Therefore, we aimed to develop an FMDV experimental model that can be handled in BSL-2 laboratories. The NanoBiT luciferase (Nano-luc) assay is a well-known assay for studying protein–protein interactions. To apply the NanoBiT split luciferase assay to the diagnosis and evaluation of FMD, we developed an inactivated HiBiT-tagged Asia1 Shamir FMDV (AS-HiBiT), a recombinant Asia1 shamir FMDV with HiBiT attached to the VP1 region of Asia1 shamir FMDV. In addition, we established LgBiT-expressing LF-BK cell lines, termed LgBit-LF-BK cells. It was confirmed that inactivated AS-HiBiT infected LgBiT-LF-BK cells and produced a luminescence signal by binding to the intracellular LgBiT of LgBiT-LF-BK cells. In addition, the luminescence signal became stronger as the number of LgBiT-LF-BK cells increased or the concentration of inactivated AS-HiBiT increased. Moreover, we confirmed that inactivated AS-HiBiT can detect seroconversion in sera positive for FMDV-neutralizing antibodies. This NanoBiT split luciferase assay system can be used for the diagnosis and evaluation of FMD and expanded to FMD-like virus models to facilitate the evaluation of FMDV vaccines and antibodies.
