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Abstract
The antigenic variation of influenza A virus hemagglutinin (HA) and neuraminidase
(NA) glycoproteins requires frequent changes in vaccine formulation. The classical
method of creating influenza virus seed strains for vaccine production is to generate
6 + 2 reassortants that contain six genes from a high-yield virus, such as A/PR/8/34
(H1N1) and the HA and NA genes of the circulating strains. The techniques currently
used are time-consuming because of the selection process required to isolate the reassortant
virus. We generated the high-yield virus A/PR/8/34 (H1N1) entirely from eight plasmids.
Its growth phenotype in embryonated chicken eggs was equivalent to that of the wild-type
virus. By using this DNA-based cotransfection technique, we generated 6 + 2 reassortants
that had the antigenic determinants of the influenza virus strains A/New Caledonia/20/99
(H1N1), A/Panama/2007/99 (H3N2), A/teal/HK/W312 (H6N1), and A/quail/HK/G1/97 (H9N2).
Our findings demonstrate that the eight-plasmid system allows the rapid and reproducible
generation of reassortant influenza A viruses for use in the manufacture of vaccines.
Copyright 2002 Elsevier Science Ltd.