Establishment of a new method based on NA activity for detecting virus titers of cell culture-based influenza vaccines
10.3760/cma.j.cn112309-20191213-00422
- VernacularTitle:基于NA活性检测细胞基质流感疫苗病毒滴度新方法的建立
- Author:
Hui ZHAO
1
;
Jianfeng WANG
;
Zhifang YING
;
Kangwei XU
;
Juan LI
;
Changgui LI
Author Information
1. 中国食品药品检定研究院呼吸道病毒疫苗室，北京 102629
- From:
Chinese Journal of Microbiology and Immunology
2020;40(4):295-299
- CountryChina
- Language:Chinese
-
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.
