两种低pH孵放法灭活静注人免疫球蛋白中脂包膜病毒效果比较
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摘要
目的比较两种低pH孵放法对静注人免疫球蛋白(intravenous immunoglobulins,IVIG)中脂包膜病毒的灭活效果。方法 IVIG中分别加入两种核酸类型的脂包膜病毒水疱性口炎病毒(vesicular stomatitis virus,VSV)及伪狂犬病毒(pseudorabies virus,PRV)作为指示病毒,以经典工艺(23～25℃灭活21 d)及新工艺[(37±0. 5)℃灭活9 h]低pH(pH为3. 8～4. 4)孵放法进行病毒灭活,分别检测不同灭活时间的病毒滴度,比较灭活效果。结果经典工艺低pH孵放法对VSV灭活效果≥5. 88 logTCID_(50)/0. 1 mL,对PRV的灭活效果≥6. 00 logTCID_(50)/0. 1 mL,且灭活的样品在敏感细胞盲传3代无细胞病变。新工艺低pH孵放法对VSV灭活效果≥4. 62 logTCID_(50)/0. 1 mL,对PRV的灭活效果≥6. 12 logTCID_(50)/0. 1 mL。结论两种低p H孵放法对指示病毒的灭活能力均符合国家标准,但经典工艺对两种指示病毒的灭活效果好且灭活彻底。
Objective To compare two low pH incubation methods for inactivation of lipid enveloped virus in intravenous immunoglobulins(IVIG). Methods Lipid enveloped virus of two nucleic acid types,vesicular stomatitis virus(VSV,RNA virus) and pseudorabies virus(PRV,DNA virus),were used as indicator virus and added to IVIG respectively,and inactivated by classical(23 ～ 25 ℃ for 21 d)and novel[(37 ± 0. 5)℃ for 9 h]low pH(3. 8 ～ 4. 4) incubation methods separately. The virus titers after inactivation for various time durations were determined to compare the inactivation effects. Results The titer of VSV after inactivation by classical low pH incubation method decreased by not less than 5. 88 logTCID_(50)/0. 1 mL,while that of PRV by not less than 6. 00 logTCID_(50)/0. 1 mL,and no CPE was found in the inactivated virus after three blind passages in sensitive cells. However,the VSV titer after inactivation by novel low pH incubation method decreased by not less than 4. 62 logTCID_(50)/0. 1 mL,while that of PRV by not less than 6. 12 logTCID_(50)/0. 1 mL. Conclusion The inactivation ability of two low p H incubation methods met the national standard.However,the classical method was more effective to inactive the two indicator viruses.
Objective To compare two low pH incubation methods for inactivation of lipid enveloped virus in intravenous immunoglobulins(IVIG). Methods Lipid enveloped virus of two nucleic acid types,vesicular stomatitis virus(VSV,RNA virus) and pseudorabies virus(PRV,DNA virus),were used as indicator virus and added to IVIG respectively,and inactivated by classical(23 ～ 25 ℃ for 21 d)and novel[(37 ± 0. 5)℃ for 9 h]low pH(3. 8 ～ 4. 4) incubation methods separately. The virus titers after inactivation for various time durations were determined to compare the inactivation effects. Results The titer of VSV after inactivation by classical low pH incubation method decreased by not less than 5. 88 logTCID_(50)/0. 1 mL,while that of PRV by not less than 6. 00 logTCID_(50)/0. 1 mL,and no CPE was found in the inactivated virus after three blind passages in sensitive cells. However,the VSV titer after inactivation by novel low pH incubation method decreased by not less than 4. 62 logTCID_(50)/0. 1 mL,while that of PRV by not less than 6. 12 logTCID_(50)/0. 1 mL. Conclusion The inactivation ability of two low p H incubation methods met the national standard.However,the classical method was more effective to inactive the two indicator viruses.
引文
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[2]YE D Z,WANG C F.Clinical efficiency of high-dose methylprednisolone combined with gamma globulin in the treatment of hand-foot-and-mouth disease[J].Clin Med Res Practic,2018,3(10):98-99.(in Chinese)叶德志,王成芳.大剂量甲泼尼龙联合丙种球蛋白治疗手足口病临床疗效[J].临床医学研究与实践,2018,3(10):98-99.
[3]BALLOW M C.84-Immunoglobulin therapy:replacement and immunomodulation//Clinical immunology[M].Amsterdam:Elsevier,2019:1143-1153.
[4]SAEEDIAN M,RANDHAWA I.Immunoglobulin replacement therapy:a twenty-year review and current update[J].Int Arch Allergy Immunol,2014,164(2):151-166.
[5]LANE R S.Non-A non-B hepatitis from intravenous immunoglobulin[J].Lancet,1983,2(8360):974-975.
[6]MATHEW A M,MUN A B,BALAKRISHNAN A.Ultraviolet inactivation of chikungunya virus[J].Intervirology,2018,61(1):36-41.
[7]BURNUM-JOHNSON K E,KYLE J E,EISFELD A J,et al.MPLEx:a method for simultaneous pathogen inactivation and extraction of samples for multi-omics profiling[J].The Analyst,2017,142(3):442-448.
[8]KEIL SD,BOWEN R,MARSCHNER S.Inactivation of Middle East respiratory syndrome coronavirus(MERS-CoV)in plasma products using a riboflavin-based and ultraviolet light-based photochemical treatment[J].Transfusion,2016,56(12):2948-2952.
[9]MARKUS E,UTE G,WIEBKE H,et al.Inactivation of Ebola virus and Middle East respiratory syndrome coronavirus in platelet concentrates and plasma by ultraviolet C light and methylene blue plus visible light,respectively[J].Transfusion,2018,58(9):2202-2207
[10]KUHNEL D,MULLER S,PICHOTTA A,et al.Inactivation of Zika virus by solvent/detergent treatment of human plasma and other plasma-derived products and pasteurization of human serum albumin[J].Transfusion,2017,57(3pt2):802-810.
[11]World Health Organization.Guidelines on viral inactivation and removal procedures intended to assure the viral safetyof human blood plasma products[J].WHO Technical Report,2004(924):219-224.
[12]国家食品药品监督管理局.动物源性医疗器械产品注册申报资料撰写指导原则[S].2012-05-19.