Screening and application of a verification method for influenza virus inactivation in MDCK cells
10.13200/j.cnki.cjb.004412
- VernacularTitle:流感病毒在MDCK细胞上灭活验证方法的筛选及应用
- Author:
LV Yuan
- Publication Type:Journal Article
- Keywords:
Influenza virus;
Influenza vaccine;
MDCK cells;
Virus inactivation
- From:
Chinese Journal of Biologicals
2025;28(02):197-203
- CountryChina
- Language:Chinese
-
Abstract:
Objective To screen a verification method for influenza virus inactivation in MDCK cells, and apply it to the sample detection before and after influenza virus inactivation, so as to provide a reliable method for the quality detection of influenza vaccines. Methods Four types of influenza viruses adapted to MDCK cells, B/Yamagata(BY), B/Victoria(BV),H1N1 and H3N2, were diluted to 10, 1, 0. 1, 0. 01, 0. 001, and 0. 000 1 lgCCID_(50)/mL separately, which were then inoculated into MDCK cells and cultured by changing the medium(discarding after adsorption for 1. 5 h and adding virus culture medium, passaging every 3 d for a total of 3 generations), rehydration method(adding virus culture medium directly after adsorption for 1. 5 h, passaging every 3 d for a total of 3 generations), and method in European Pharmacopoeia(passaging every 4 d for a total of 3 generations). The cytopathic effect(CPE) was observed every day, the virus solutions of different passages were harvested, and the hemagglutinin(HA) titers were detected to determine the optimum inactivation verification method. Finally, the optimum inactivation verification method was used to detect samples before and after inactivation of influenza virus. Results Influenza virus could be detected at the titer of 0. 1-1 lgCCID_(50)/mL by both changing medium method and rehydration method, while influenza virus could be detected in the titer range of 1-10 lgCCID_(50)/mL by test protocol in European Pharmacopoeia, and the titer of HA detected by rehydration method was higher than those by the other two methods. Therefore, rehydration method was determined as the optimum inactivation verification method. The samples detected by rehydration method were all positive before influenza virus inactivation and negative after virus inactivation.Conclusion Rehydration method is the optimum method to verify the influenza virus inactivation in MDCK cells, which can be used to detect the quality of influenza vaccines.
