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Error-prone pcr-based mutagenesis strategy for rapidly generating high-yield influenza vaccine candidates.

Virology (2015-04-23)
Jianqiang Ye, Feng Wen, Yifei Xu, Nan Zhao, Liping Long, Hailiang Sun, Jialiang Yang, Jim Cooley, G Todd Pharr, Richard Webby, Xiu-Feng Wan
ABSTRACT

Vaccination is the primary strategy for the prevention and control of influenza outbreaks. However, the manufacture of influenza vaccine requires a high-yield seed strain, and the conventional methods for generating such strains are time consuming. In this study, we developed a novel method to rapidly generate high-yield candidate vaccine strains by integrating error-prone PCR, site-directed mutagenesis strategies, and reverse genetics. We used this method to generate seed strains for the influenza A(H1N1)pdm09 virus and produced six high-yield candidate strains. We used a mouse model to assess the efficacy of two of the six candidate strains as a vaccine seed virus: both strains provided complete protection in mice against lethal challenge, thus validating our method. Results confirmed that the efficacy of these candidate vaccine seed strains was not affected by the yield-optimization procedure.

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