Abstract
The antigen variability of the infectious bronchitis virus (IBV) has hindered vaccine effectiveness and perpetuated its epidemic. We engineered a rapid attenuation method for IBV variants. The strategy involves creating the rH-CPDF7 backbone by recoding a segment of the H120 nonstructural protein (NSP) genome via codon pair deoptimization (CPD), facilitating S gene integration from IBV variants via transformation-associated recombination (TAR) cloning. These recombinant strains exhibited even lower pathogenicity, indicating the effectiveness of CPDF7 in reducing virulence. Importantly, the rH-CPDF7 backbone demonstrated versatility, being applicable to the development of attenuated strains for IBV variants, including the QX-type, TW-type, and GVI-type strains (different genotypes). In conclusion, our method allows for the rapid development of attenuated strains by integrating the S gene of IBV variants into the rH-CPDF7 backbone. These recombinant strains can elicit a strong immune response and provide effective protection against homologous challenges. This strategy is crucial for developing live-attenuated vaccines against emerging IBV strains.