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水痘-带状疱疹病毒快速微量中和试验的建立与初步应用 被引量:1

Establishment and preliminary application of a rapid microneutralization test for screening varicella-zoster virus special hyperimmune globulin in plasma donations
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摘要 目的 建立水痘-带状疱疹病毒(VZV)的快速微量中和试验筛查技术,用于高免原料血浆的筛选和VZV特异性免疫球蛋白(VZIG)的制备。方法 参考美国FDA批准的VZIG制品,设立原料血浆筛选滴度的下限;通过病毒滴度优化,设立快速微量中和试验筛查体系中的病毒使用滴度;建立筛选体系并引入VZIG国际标准品作为质量控制,对1∶2~1∶256倍比稀释的各滴度的单人份和混合原料血浆进行测定,同时利用抗VZV中和抗体ELISA检测试剂盒Vacc Zyme复核快速微量中和筛选结果,对其灵敏度进行分析。结果快速微量中和筛选试验中原料血浆的最低筛选下限为1∶16(VZV中和抗体滴度不低于0.4 U/ml);筛选体系病毒使用滴度为1500 PFU/ml;20份单采血浆中有8人份的滴度≥1∶16,混合血浆中无符合筛选标准的样本;ELISA的复核结果与快速微量中和试验呈良好的直线关系(r=0.895 24,P〈0.0001)。结论 快速微量中和试验的灵敏性、通量和可操作性等,适于VZV高免原料血浆的筛选,解决了VZIG制备中原料血浆筛选的关键技术问题。 Objective To promote the progress in varicella-zoster virus (VZV) immunoglobulin preparation, a rapid mieroneutralization test (RMNt) was set up for screening plasmapheresis donations with high titers of special neutralizing antibodies to VZV. Methods With reference to the VZV imnmnoglobulin (VZIG) preparation standard of FDA and VZIG international unit (IU) , a screening standard was formulated; the amount of virus was analyzed to determine the optimal conditions for RMNt; screening technology was established and the IU was introduced as quality control; twenty samples of apheresis plasma and fifteen samples of pooled plasma were diluted at 1 : 2 to 1 : 256 and tested by RMNt respectively; and the sensitivity of RMNt was also analyzed by the commercial ELISA kit. Results Plasma samples that were diluted at 1 : 16 and had a titer more than 0.4 U/ml could be used in the production of VZIG. 1500 PFU/ml titers of virus in RMNt pro- duced readable results in plasma screening. Eight apheresis plasma samples tested met the screening standard; but none of the pooled samples tested positive. RMNt had a good linear relationship with ELISA ( r = 0. 895 24 ,P 〈 0. 0001 ). Conclu- sion The sensitivity, throughput and operability of the established RMNt can be used in the screening of plasma donations as key techniques for the production of VZIG.
作者 王卓 吕茂民 赵雄 马玉媛 赵媛媛 章金刚 WANG Zhuo, LU Mao-min , ZHAO Xiong, MA Yu-yuan , ZHAO Yuan-yuan, ZHANG Jin-gang ( 1. Institute of Transfusion Medicine, Academy of Military Medical Sciences, Beijing 100850, China; 2. Department of Clinical Laboratory, No. 457 Hospital of PLA, Wuhan 430012, China;3. Outpatient Clinic, Logistics College of PLA, Bei- jing 100858, China)
出处 《军事医学》 CAS CSCD 北大核心 2015年第3期179-183,共5页 Military Medical Sciences
基金 国家863计划资助项目(2012AA021902)
关键词 水痘-带状疱疹病毒 免疫球蛋白类 快速微量中和试验 血浆 varicella-zoster virus immunoglobulins rapid microneutralization test plasma
作者简介 王卓,女,博士,研究方向:血液制品 吕茂民,E-mail:maominlv@126.com [通讯作者]:章金刚,E-mail:zhangjg@bmi.ac.cn;
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