低pH胃蛋白酶-甲苯消化法灭活破伤风免疫血浆中脂包膜病毒效果验证  

Verification of inactivation efficiency of lipid-enveloped viruses in immune plasma against tetanus by pepsin-toluene digestion at low pH value

作　　者：王剑锋[1] 英志芳[1] 李长贵[1] 

机构地区：[1]中国食品药品检定研究院病毒三室,北京100050

出　　处：《中国生物制品学杂志》2012年第9期1214-1215,1219,共3页Chinese Journal of Biologicals

摘　　要：目的验证低pH条件下胃蛋白酶-甲苯消化法对不同核酸类型的脂包膜病毒的灭活效果。方法以水疱性口炎病毒(Vesicular stomatitis virus,VSV)作为RNA指示病毒,以伪狂犬病病毒(Pseudorabies virus,PRV)作为DNA指示病毒,将破伤风免疫血浆样品经低pH(3.2±0.2)、0.2%甲苯、6～12活力单位胃蛋白酶处理后各取27 ml,分别加入3 ml指示病毒(9∶1),在(30.0±1)℃水浴中分别振摇10、30、60、90 min灭活病毒。以Vero细胞、PK-15细胞为基质,采用96孔细胞病变法检测残余病毒滴度,验证病毒灭活效果。结果破伤风免疫血浆样品经低pH胃蛋白酶及甲苯消化处理90 min后,VSV和PRV两种指示病毒的灭活效果分别为3.12～3.88和5.25～5.38 logTCID50/0.1 ml。结论采用低pH胃蛋白酶-甲苯消化法对破伤风免疫血浆中PRV灭活效果较好,而对VSV灭活效果有待提高。Objective To verify the inactivation efficiency of lipid-enveloped viruses of various nucleic acid patterns by pepsin-toluene digestion at low pH value.Methods Pseudorabies virus（PRV） and vasicular stomatitis virus（VSV） were used as indicators for DNA and RNA lipid-enveloped viruses,respectively.The immune plasma against tetanus was treated with toluene（0.2%） and pepsin（6 ～ 12 U） at pH（3.2 ± 0.2）,from which 27 ml was taken,added with 3 ml of indicator（9 ∶ 1）,then incubated in（30 ± 1）℃ water bath for 10,30,60 and 90 min in shaking.Vero and PK-15 cells were used substrates for measuring the residual virus titer by CPE method on 96-well microplate,based on which the virus inactivation efficiency was verified.Results The titers of VSV and PRV after pepsin-toluene digestion for go min decreased by not less than 3.12 ～ 3.88 and not less than 5.25 ～ 5.38 logTCID50 / 0.1 ml respectively.Conclusion PRV in immune plasma against tetanus was inactivated effectively by pepsin-toluene digestion.However,the inactivation efficiency of VSV was to be further improved.

关 键 词：破伤风免疫血浆 脂包膜病毒 胃蛋白酶 甲苯 病毒灭活 

分 类 号：R378.81[医药卫生—病原生物学] R392-33[医药卫生—基础医学]