Establishment of a new method based on NA activity for detecting virus titers of cell culture-based influenza vaccines / 中华微生物学和免疫学杂志
Chinese Journal of Microbiology and Immunology
; (12): 295-299, 2020.
Article
em Zh
| WPRIM
| ID: wpr-871277
Biblioteca responsável:
WPRO
ABSTRACT
Objective:To develop a new method based on neuraminidase (NA) activity for detec-ting virus titers of cell culture-based influenza vaccines and preliminarily analyze its application.Methods:Reaction conditions including the substrate concentration for enzymatic reaction, stop solution, the number of initially infected target cells and cell lysis buffer were optimized. The titers of cell culture-based influenza vaccine strains were detected by the established method and the results were compared with those by the traditional viral titration test.Results:The optimal substrate concentration for enzymatic reaction was 25 μmol/L, and the optimal stop solution was 0.2 mol/L Na 2CO 3. In the detection of NA activity in infected cells, the maximum relative fluorescence value was obtained by infecting 4×10 4 cells/well with influenza virus for 48 h and using 0.5% TritonX-100 for lysis. The developed method showed no significant differences with the traditional virus titration test in detecting the titers of four batches of influenza vaccine virus strains ( P>0.05), indicating that the two methods had a good consistency. Conclusions:This study established a new method based on NA activity to detect virus titers of cell culture-based influenza vaccines. The method could be used for the detection of virus strains used in the production of cell cultured-based influenza vaccines.
Texto completo:
1
Base de dados:
WPRIM
Idioma:
Zh
Revista:
Chinese Journal of Microbiology and Immunology
Ano de publicação:
2020
Tipo de documento:
Article