Study On Virus Inactivation In DBJ By UV Irradiation

Objective:This paper mainly selected the virus physical and chemical properties are representative(virus size,nucleic acid type and whether there is an envelope),and should at least include a virus that has obvious resistance to physical and/or chemical treatment,so chose Pseudorabies Virus(PRV),Vesicular Stomatitis Virus(VSV),Sindbis virus(SINV)as lipid envelope indicator virus,Porcine parvovirus(PPV)as non-The lipid envelope indicates the virus,and the inactivation process is verified.According to the requirements of the"Technical Method for the Removal/Inactivation of Blood Products and the Guiding Principles for Validation",the titer reduction should be?4 logs.Research Methods:In this paper,the doses of ultraviolet irradiation were set to 0J/m~2,300J/m~2,450J/m~2,600J/m~2,750J/m~2,and 900J/m~2.The fibrinogen/thrombin and virus were mixed in a volume ratio of 9:1.After UV irradiation,the virus titer before and after irradiation was measured by a cytopathic method,and the virus titer was calculated by the Reed-Muench method.The protein content of fibrinogen before and after ultraviolet irradiation was measured by ultraviolet spectrophotometry,and the thrombin titer before and after ultraviolet irradiation was measured according to the method for measuring the titer of thrombin lyophilized powder in Chinese Pharmacopoeia 2015 edition,and the ultraviolet irradiation was evaluated by SDS-PAGE.Before and after the change in the protein structure of fibrinogen and thrombin.In this experiment,the UV irradiation dose in the experiment was applied to a flow-type UV inactivation device to verify the inactivation effect of the flow-type UV inactivation device.Results:UV doses of 900 J/m~2 and 600 J/m~2 can effectively inactivate porcine parvovirus,pseudorabies virus,Sindbis virus and vesicular stomatitis in fibrinogen and thrombin,respectively,resulting in a decrease in virus titer.?4.00 logs.There was no significant difference in the concentration of fibrinogen before and after UV irradiation at a dose of 900J/m~2 and 600 J/m~2 UV irradiation.There was a significant difference in the titers of thrombin,but the thrombin activity was still specified in the state.Content range(85%to 115%).Before and after irradiation,the protein band of fibrinogen and thrombin and the band before irradiation were in the same position,and the structure of the protein did not change.At a UV irradiation dose of 300 J/m~2,the flow-type UV inactivation device can also reduce the inactivation of pseudorabies virus in fibrinogen by?4.00 logs,thus meeting the requirements for virus inactivation.Conclusions:UV irradiation doses of 900 J/m~2 and 600 J/m~2 can inactivate viral titers of fibrinogen and thrombin in both lipid-enveloped and non-lipid-enveloped viruses by?4 logs,indicating that UV irradiation can be effective.Inactivated lipid-enveloped viruses and non-lipid enveloped viruses have a stable and effective inactivation effect.The protein content and structure of fibrinogen and thrombin did not change significantly before and after UV irradiation.