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Isolation of Fidelity Variants of RNA Viruses and Characterization of Virus Mutation Frequency
Published on: June 16, 2011
Characterization of a Virus Rescued from a Full-Length Infectious Clone Derived from the Type A Foot-and-Mouth
Jae Young Kim1, Sun Young Park1, Gyeongmin Lee1
1Center for Foot-and-Mouth Disease Vaccine Research, Animal and Plant Quarantine Agency, 177 Hyeoksin 8-ro, Gimcheon-si 39660, Gyeongsangbuk-do, Republic of Korea.
Abstract:
Foot-and-mouth disease (FMD), a vesicular disease, causes lesions in the mouth, nose, teats, and feet of cloven-hoofed animals. Vaccination remains the most effective method to prevent FMD outbreaks. Since 2010, South Korea has implemented nationwide vaccination and developed multiple domestic vaccine strains to achieve vaccine self-sufficiency. Here, we aimed to construct an infectious clone using the A/SKR/Yeoncheon/2017 virus, which exhibits the highest antigen productivity among previously developed vaccine strains. An infectious clone was constructed based on the A/Yeoncheon/SKR/2017 virus isolated during an FMD outbreak in Korea in 2017. The viral genome was amplified in two fragments and assembled into a full-length clone, from which infectious recombinant virus was successfully rescued. The rescued virus was confirmed via serotyping and transmission electron microscopy to exhibit canonical 25-30 nm icosahedral morphology. Under optimized culture conditions using suspension-adapted BHK-21 cells (multiplicity of infection 0.001; 12 h post-infection), the recombinant virus achieved titers of 108 TCID50/mL and produced 6.2 μg/mL of 146S antigen, comparable to its parental counterpart. The experimental vaccine formulated with the rescued virus (15 μg/dose), 1% saponin, 1% aluminum hydroxide gel, and ISA 206 VG, induced protective immunity in eight-week-old pigs, with vaccinated animals exhibiting no clinical signs following homologous challenge. To our knowledge, this study represents the first successful construction of an infectious clone derived from a field-isolated serotype A FMDV in South Korea. In the future, this A/SKR/Yeoncheon/2017 infectious clone can serve as a platform backbone for the rapid development of next-generation, high-yield vaccine seed strains through targeted epitope exchange.
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