用于SARS-CoV-2疫苗免疫后血浆筛选的不同ELISA试剂盒适用性评估  

Applicability evaluation of different ELISA kits for SARS-CoV-2 antibody screening of post-immunization plasma

作　　者：周志军 王倩 彭焱 岳胜兰 熊伟 纪德铭 冯璐 李娟 李策生 胡勇 李陶敬 ZHOU Zhijun;WANG Qian;PENG Yan;YUE Shenglan;XIONG Wei;JI Deming;FENG Lu;LI Juan;LI Cesheng;HU Yong;LI Taojing(Sinopharm Wuhan Plasma-derived Biotherapies Co.,Ltd.,Wuhan 430207,Hubei Province,China;不详)

机构地区：[1]国药集团武汉血液制品有限公司,湖北武汉430207 [2]中国生物技术股份有限公司,北京100020 [3]成都蓉生药业有限责任公司,四川成都610041

出　　处：《中国生物制品学杂志》2023年第12期1476-1482,1490,共8页Chinese Journal of Biologicals

基　　金：国家科技部重点专项(2020YFC0841800)。

摘　　要：目的通过比较3个厂家4种SARS-CoV-2 ELISA-IgG及中和抗体检测试剂盒筛选免疫后血浆的收浆合格率并比较操作便利性,确定最佳原料血浆筛选用试剂盒。方法以从现有SARS-CoV-2疫苗免疫后血浆中筛选的不同ELISA稀释倍数档位单份血浆为原料,用不同厂家SARS-CoV-2 ELISA-IgG及中和抗体检测试剂盒进行检测,按照各厂家效价分档位进行混合。混合血浆进行假病毒中和试验,根据中和抗体效价确定不同试剂盒的收浆标准,并计算各自收浆合格率;根据收浆标准确定单份血浆筛选的稀释方式,比较操作便利性;综合判断试剂盒的适用性。结果以内控参考品B2为收浆标准,厂家B ELISA-IgG抗体检测试剂盒在1108份WT>16的血浆中收浆合格率为82.1%,高于现用厂家A ELISA-IgG抗体检测试剂盒(59.8%)。以厂家B中和抗体检测试剂盒的50 IU/mL为收浆标准,该试剂盒在387份WT16~64的血浆中收浆合格率为66.4%,高于厂家A ELISA-IgG抗体检测试剂盒的收浆合格率(42.1%)。以厂家C中和抗体检测试剂盒100 IU/mL为收浆标准,该试剂盒在536份WT16~64的血浆中收浆合格率为42.5%,低于厂家A ELISA-IgG抗体检测试剂盒的收浆合格率(50.2%)。厂家C中和抗体检测试剂盒操作步骤最多,反应时间最长;厂家B ELISA-IgG抗体检测试剂盒反应时间最短,对于WT16~64的样品无需孔外稀释,而厂家A需孔外稀释。4种试剂盒的收浆合格率差异主要发生在WT16~64的样品中,其收浆合格率在该区间排序依次为:厂家B ELISA-IgG抗体检测试剂盒>厂家B中和抗体检测试剂盒>厂家A ELISA-IgG抗体检测试剂盒>厂家C中和抗体检测试剂盒;操作便利性排序为:厂家B ELISA-IgG抗体检测试剂盒>厂家B中和抗体检测试剂盒>厂家A ELISAIgG抗体检测试剂盒>厂家C中和抗体检测试剂盒。结论从收浆合格率及操作便利性综合考量试剂盒的适用性,以厂家B ELISA-IgG抗体检测试剂盒最佳。Objective To determine the optimal plasma screening kit for raw materials by comparing the qualified rate of plasma collection and operation convenience of four SARS-CoV-2 ELISA-IgG and neutralizing antibody kits from three manufacturers.MethodsSingle plasma samples with different ELISA dilution multiples screened from existing plasma immunized with SARS-CoV-2 vaccine were used as raw materials,which were detected by using SARS-CoV-2 ELISA-IgG and neutralizing antibody kits from different manufacturers.Plasma samples were mixed according to the potency level of each manufacturer,and pseudovirus neutralization test was carried out on pooled plasma.According to the potency of neutralizing antibody,the plasma collection standard of different kits were determined,and the qualified rate of plasma collection was calculated respectively.The dilution method of single plasma sample screening was determined in regard of the plasma collection standard,and the operation convenience was compared;The applicability of the kits were evaluated comprehensively.ResultsThe qualified rate of ELISA-IgG antibody kit of manufacturer B in 1108 WT>16 plasma samples was 82.1%with internal control reference B2 as the plasma collection standard,which was higher than that of the current ELISA-IgG antibody kit of manufacturer A(59.8%).With the level of 50 IU/mL tested by neutralizing antibody kit of manufacturer B as the standard of plasma collection,the qualified rate of plasma collection of this kit in 387 WT16~64 samples was 66.4%,higher than that of ELISA-IgG kit of manufacturer A(42.1%).While with the level of 100 IU/mL tested by neutralizing antibody kit of manufacturer C as the standard of plasma collection,the qualified rate of plasma collection of this kit in 536 WT16~64 samples was 42.5%,lower than that of ELISA-IgG kit of manufacturer A(50.2%).Neutralizing antibody kit of manufacturer C had the most operation steps and the longest reaction time;ELISA-IgG kit of manufacturer B had the shortest reaction time,and required no out-of-hol

关 键 词：SARS-CoV-2 ELISA-IgG 中和抗体 稀释倍数 免疫后血浆 收浆合格率 

分 类 号：R392-33[医药卫生—免疫学]