Implementation of new approaches for generating conventional reassortants for live attenuated influenza vaccine based on Russian master donor viruses

摘要: Cold-adapted influenza strains A/Leningrad/134/17/57 (H2N2) and B/USSR/60/69, originally developed in Russia, have been reliable master donors of attenuation for preparing live attenuated vaccines (LAIV). The classical strategy generating LAIV reassortants is robust, but has some disadvantages. generation requires at least 3 passages under selective conditions after co-infection; each these takes six days. Screening the a genomic composition traditionally starts second limiting dilution cloning procedure, number suitable limited. We new approach to shorten process seed viruses. Introducing genotyping by pyrosequencing monitoring sequence integrity surface antigens starting first passage allowed specific selection next procedure also eliminate one group vaccine candidate generation. Homogeneity analysis confirmed that reducing didn’t affect quality Finally, two-way hemagglutination inhibition test, implemented all final viruses, any amino acid substitutions acquired during egg propagation antigenicity vaccine. Our reduces time required generate was used seasonal LAIVs candidates 2012/2013, 2014/2015, 2015/2016 seasons more rapidly.