羧甲基壳聚糖病毒灭活/去除工艺验证  

Verification of inactivation / removal procedure for carboxymethyl chitosan

作　　者：杨倩[1] 宋战昀[1] 张旭光[1] 李敏思[1] 王向辉[1] 隋佳辰 张健[1,2] 郑言[1] 魏春艳[1] 王振国[1] 

机构地区：[1]吉林出入境检验检疫局检验检疫技术中心,吉林长春130062 [2]长春生物制品研究所有限责任公司,吉林长春130062

出　　处：《中国生物制品学杂志》2016年第5期533-537,共5页Chinese Journal of Biologicals

基　　金：国家自然科学基金项目(31001065);国家质检总局科技计划项目(2013IK033)

摘　　要：目的对羧甲基壳聚糖病毒灭活/去除工艺进行验证。方法模拟生产条件,选择猪细小病毒(porcine parvovirus,PPV)、伪狂犬病病毒(pseudo rabies virus,PRV)、鸭肝炎病毒Ⅰ型(duck hepatitis virus-Ⅰ,DHV-Ⅰ)、牛滤过性腹泻病毒(bovine viral diarrhea virus,BVDV)作为指示病毒,采用高温碱化和醇洗灭活/去除病毒,Karber法计算病毒滴度,荧光PCR检测病毒灭活/去除效果。结果 3批甲壳素经碱化反应处理,PRV、PPV、BVDV、DHV-Ⅰ的平均灭活对数值分别为≥6.736、≥6.597、≥6.138和≥5.806 log TCID50/0.1 ml;3批羧甲基壳聚糖经醇洗处理,PRV、PPV、BVDV、DHV-Ⅰ的平均灭活对数值分别为≥6.763、≥6.569、≥6.167和≥5.587 log TCID50/0.1 ml。碱化反应与醇洗处理前感染PRV、PPV、BVDV、DHV-Ⅰ样品荧光PCR检测平均Ct值分别为23.465、23.387,24.873、24.706,26.887、25.376,27.386、24.629,处理后样品的荧光PCR检测平均Ct值均>40。结论羧甲基壳聚糖经高温碱化和醇洗反应处理均能有效灭活病毒,保证了产品的安全性。Objective To verify the inactivation / removal of procedure for carboxymethyl chitosan.Methods Under the mimic production condition,the porcine parvovirus（PPV）,pseudorabies virus（PRV）,duck hepatitis virus-Ⅰ（DHV-Ⅰ）and bovine viral diarrhea virus（BVDV）were used as indicators for verification of virus inactivation / removal procedure of carboxymethyl chitosan by high temperature alkalization and alcoholization.The virus titer was calculated by Karber method,while the virus inactivation / removal efficacy was evaluated by fluorescent PCR.Results The average logarithm inactivation value（LIV）of PRV,PPV,BVDV and DHV-Ⅰin three batches of chitosan after alkalization were not less than6.736,not less than 6.597,not less than 6.138 and not less than 5.806 log TCID_（50）/ 0.1 ml,while those in three batches of carboxymethyl chitosan after alcoholization were not less than 6.763,not less than 6.569,not less than 6.167 and not less than 5.587 log TCID_（50）/ 0.1 ml,respectively.The Ct values of samples infected with PRV,PPV,BVDV and DHV-Ⅰbefore alkalization were 23.465,24.873,26.887 and 27.386,while those before alcoholization were 23.387,24.706,25.376 and 24.629,respectively.However,all the Ct values after treatment were more than 40.Conclusion Both high temperature alkalization and alcoholization may inactivate the viruses in carboxymethyl chitosan effectively and ensure the safety of the product.

关 键 词：甲壳素 羧甲基壳聚糖 病原体灭活/去除 验证 

分 类 号：R392.33[医药卫生—免疫学]