短波紫外线对人凝血因子Ⅷ中猪细小病毒灭活的研究

摘要

Objective To investigate the effect of short-wave ultraviolet-C (UV-C) on inactivating porcine parvovirus (PPV) in human coagulation factor Ⅷ (FⅧ).Methods PPV was mixed with the FⅧ intermediate as an indicator virus.The FⅧ intermediates containing PPVs were inactivated using UVivatec instrument with 200,300,and 400 J/m2 UV-C.The FⅧ s inactivated by UV-C were inoculated in ST cells and cultured.PPV was detected by cytopathic effect (CPE) method and the virus titer was calculated by Reed-Muench method.The samples without CPE were cultured 3 blind passages to verify the inactivation effect.At the same time,the FⅧ activities before and after UV-C inactivation were detected.Results The PPV titers in FⅧ all decreased no less than 4.62 lg 50％ tissue culture infective dose per 0.1 ml after UV-C inactivation with 3 exposure doses.PPVs were not detected in the samples without CPE cultured to 3rd blind passage.The activities of FⅧ inactivated by UV-C did not decrease significantly,and each quality index met the requirements of pharmacopeia.Conclusion UV-C can inactivate nonenveloped virus PPV,and has no influence on the FⅧ activity.%目的 研究短波紫外线(short-wave ultraviolet-C,UV-C)对人凝血因子Ⅷ(human coagulationfactor Ⅷ,FⅧ)中猪细小病毒(porcine parvovirus,PPV)的灭活的效果.方法 将PPV作为指示病毒与FⅧ中间品混合,用紫外灭活仪UVivatec分别以200、300和400 J/m2 UV-C进行灭活.将UV-C灭活的FⅧ接种ST细胞并进行培养,采用细胞病变(cytopathic effect,CPE)法检测病毒,并以Reed-Muench法计算病毒滴度.对所有UV-C灭活后未出现CPE的样品盲传3代,验证灭活效果;同时对UV-C灭活前后的FⅧ活性进行检测.结果 3个照射剂量的UV-C灭活后,FⅧ中的PPV滴度降低均不低于每0.1 ml4.62lg半数组织培养感染剂量,所有未出现CPE的样品盲传至第3代均未检到病毒.UV-C灭活的FⅧ的活性无明显降低,且各项质量指标均符合药典的要求.结论 UV-C可有效灭活无包膜病毒PPV,且对FⅧ活性无明显影响.