Screening of the high yield influenza B virus on MDCK cell and cloning of its whole genome

Abstract

Influenza virus may cause pandemic in the world. Thus influenza vaccine should be produced timely and effectively. There are many advantages in producing influenza vaccine with mammalian cell culture rather than with chicken embryos. But it is difficult to obtain high titer of the virus on MDCK cell. We have passaged B/jingke/96/01 influenza virus strain on MDCK cells for 15 passages, the PFU reached from the first passage 8×10 4, to the 15th passage 2×10 9, the TCID 50/ml was 6.5 of the first passage to 8.0 of the 15th passage. Not only B/jingke/96/01 has high productive characteristics in MDCK cells, but also has a tendency of increasing. We designed two sets of universal primers to amplify the whole genome of influenza virus B/jingke/96/01. In order to produce the recombinant influenza vaccine, this strain may be reassorted with the current influenza virus strain as a master strain by reverse genetic technique.