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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 establish a rapid micro-neutralization screening test for varicella-zoster virus (VZV) for the screening of plasma-free raw materials and the preparation of VZV-specific immunoglobulin (VZV). Methods Reference to the FDA approved VZIG products, the establishment of the lower limit of the plasma screening of raw material titer; virus titer optimization set up rapid micro-neutralization test screening system virus titer; the establishment of screening system and the introduction of VZIG international standards as Quality control, single titer of 1: 2 to 1:256 dilutions of each titer and mixed raw plasma were measured, and simultaneously with the anti-VZV neutralizing antibody ELISA test kit Vacc Zyme review rapid micro-neutralization screening results, Its sensitivity is analyzed. Results The minimum screening limit of plasma was 1:16 (VZV neutralizing antibody titer was not less than 0.4 U / ml) in the rapid micro-neutralization screening test. The screening titer of virus was 1500 PFU / ml. Eight plasma samples were titrated ≥1: 16 and no plasma samples were screened. The results of ELISA were in good linear relationship with the rapid microneutralization assay (r = 0.895 24, P <0.0001). Conclusion The sensitivity, throughput and maneuverability of the rapid micro-neutralization test are suitable for the screening of VZV highly-free raw plasma, which solves the key technical problems of plasma screening of VZIG.