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

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

doi:10.3390/pathogens9030188

Cited by 13 publications

(17 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This mixture was treated by heating at 56 °C for 30 min [ 13 ]. Serum neutralization test (SNT) using a luciferase-based recombinant virus was performed according to a protocol previously described [ 14 ]. The neutralization titers of antibodies against vCSFV GPE – /HiBiT were calculated according to luciferase activity measured in the culture supernatants using Nano-Glo HiBiT lytic detection system (Promega, Madison, WI, USA).…”

Section: Methodsmentioning

confidence: 99%

Efficacy of Oral Vaccine against Classical Swine Fever in Wild Boar and Estimation of the Disease Dynamics in the Quantitative Approach

Bazarragchaa

Isoda

Kim

et al. 2021

Viruses

Self Cite

View full text Add to dashboard Cite

Classical swine fever virus (CSFV) in the wild boar population has been spreading in Japan, alongside outbreaks on pigs, since classical swine fever (CSF) reemerged in September 2018. The vaccination using oral bait vaccine was initially implemented in Gifu prefecture in March 2019. In the present study, antibodies against CSFV in wild boar were assessed in 1443 captured and dead wild boars in Gifu prefecture. After the implementation of oral vaccination, the increase of the proportion of seropositive animals and their titer in wild boars were confirmed. Quantitative analysis of antigen and antibodies against CSFV in wild boar implies potential disease diversity in the wild boar population. Animals with status in high virus replication (Ct < 30) and non- or low-immune response were confirmed and were sustained at a certain level after initial oral vaccination. Through continuous vaccination periods, the increase of seroprevalence among wild boar and the decrease of CSFV-positive animals were observed. The epidemiological analysis based on the quantitative virological outcomes could provide more information on the efficacy of oral vaccination and dynamics of CSF in the wild boar population, which will help to improve the implementation of control measures for CSF in countries such as Japan and neighboring countries.

show abstract

Section: Methodsmentioning

confidence: 99%

Efficacy of Oral Vaccine against Classical Swine Fever in Wild Boar and Estimation of the Disease Dynamics in the Quantitative Approach

Bazarragchaa

Isoda

Kim

et al. 2021

Viruses

Self Cite

View full text Add to dashboard Cite

show abstract

“…As reported, the VNT titers of the serum can be determined tentatively at 2 days post-infection (dpi) and are comparable to those obtained by conventional VNTs at 3 or 4 dpi. This system allows CSF virus growth to be easily and rapidly monitored and enabled the rapid and easy determination of the VNT titer using a luminometer, which could be a powerful tool to replace the conventional VNT as a high-throughput antibody test for CSFV infections [ 57 ].…”

Section: Antibody Detectionmentioning

confidence: 99%

Recent Advances in the Diagnosis of Classical Swine Fever and Future Perspectives

Wang

Madera

et al. 2020

Pathogens

View full text Add to dashboard Cite

Classical swine fever (CSF) is a highly contagious viral disease of pigs, including wild boar. It is regarded as one of the major problems in the pig industry as it is still endemic in many regions of the world and has the potential to cause devastating epidemics, particularly in countries free of the disease. Rapid and reliable diagnosis is of utmost importance in the control of CSF. Since clinical presentations of CSF are highly variable and may be confused with other viral diseases in pigs, laboratory diagnosis is indispensable for an unambiguous diagnosis. On an international level, well-established diagnostic tests of CSF such as virus isolation, fluorescent antibody test (FAT), antigen capture antibody enzyme-linked immunosorbent assay (ELISA), reverse-transcription polymerase chain reaction (RT-PCR), virus neutralization test (VNT), and antibody ELISA have been described in detail in the OIE Terrestrial Manual. However, improved CSF diagnostic methods or alternatives based on modern technologies have been developed in recent years. This review thus presents recent advances in the diagnosis of CSF and future perspectives.

show abstract

“…Tetsuo and colleagues developed a novel high-throughput serum neutralization test (SNT) for the serological diagnosis of infections (classical swine fever, bovine diarrhea, and borderline disease viruses) based on genetically engineered pestiviruses expressing the HiBiT peptide [ 231 ]. Swine or bovine kidney cells line were infected with recombinant viruses in serially diluted antiserum.…”

Section: Ctz-dependent Luciferase Analytical Applicationmentioning

confidence: 99%

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

Krasitskaya

Bashmakova

Frank

2020

IJMS

View full text Add to dashboard Cite

: The functioning of bioluminescent systems in most of the known marine organisms is based on the oxidation reaction of the same substrate—coelenterazine (CTZ), catalyzed by luciferase. Despite the diversity in structures and the functioning mechanisms, these enzymes can be united into a common group called CTZ-dependent luciferases. Among these, there are two sharply different types of the system organization—Ca2+-regulated photoproteins and luciferases themselves that function in accordance with the classical enzyme–substrate kinetics. Along with deep and comprehensive fundamental research on these systems, approaches and methods of their practical use as highly sensitive reporters in analytics have been developed. The research aiming at the creation of artificial luciferases and synthetic CTZ analogues with new unique properties has led to the development of new experimental analytical methods based on them. The commercial availability of many ready-to-use assay systems based on CTZ-dependent luciferases is also important when choosing them by first-time-users. The development of analytical methods based on these bioluminescent systems is currently booming. The bioluminescent systems under consideration were successfully applied in various biological research areas, which confirms them to be a powerful analytical tool. In this review, we consider the main directions, results, and achievements in research involving these luciferases.

show abstract

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

Cited by 13 publications

References 38 publications

Efficacy of Oral Vaccine against Classical Swine Fever in Wild Boar and Estimation of the Disease Dynamics in the Quantitative Approach

Efficacy of Oral Vaccine against Classical Swine Fever in Wild Boar and Estimation of the Disease Dynamics in the Quantitative Approach

Recent Advances in the Diagnosis of Classical Swine Fever and Future Perspectives

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

Contact Info

Product

Resources

About