IgG型意外抗体鉴定用试剂盒(固相凝集法)的研发  

Develop a solid-phase red cell adherence reagent kit for IgG irregular unexpected alloantibody identification

作　　者：骆宏[1] 邵媛[1] 王嘉励[1] 王贞[1] 罗广平[1] LUO Hong;SHAO Yuan;WANG Jiali;WANG Zhen;LUO Guangping(Guangzhou Blood Center,The Key Medical Laboratory of Guangzhou,Guangzhou 510095,China)

机构地区：[1]广州血液中心广州市医学(血液安全)重点实验室,广东广州510095

出　　处：《中国输血杂志》2024年第10期1158-1163,共6页Chinese Journal of Blood Transfusion

基　　金：广州市科技计划项目校企联合资助专题(2023A03J1001);广州市科技计划项目(2023A03J0552)。

摘　　要：目的研发IgG型意外抗体鉴定用试剂盒(固相凝集法)。方法琥珀酰亚胺溶液包被微孔板后,将谱细胞悬液加入微孔板中制得红细胞单层,随后用1×PBS(pH7.40)裂解红细胞单层形成细胞膜单层;将低温干燥保护液加至细胞膜单层表面,4℃干燥后制成试剂盒反应板;定期用制备的试剂盒测试低效价IgG抗-E和抗-Fy a,以评估试剂盒稳定性;对72例意外抗体阳性血浆标本和152例正常血浆标本采用研发的试剂盒和微柱凝胶卡进行抗体鉴定试验,计算灵敏度和特异度指标,采用卡方检验分析,以评估试剂盒的检测性能。结果琥珀酰亚胺包被的微板能够较好的固定裂解后的红细胞膜;低温干燥保护液对膜单层能够形成较好的保护作用;在各个观察点内,低效价的IgG抗-E和抗-Fy a均能被准确检出;与微柱凝胶卡相比,72例意外抗体阳性的标本,固相法试剂盒正确检出70例,同时纠正1例凝胶卡漏检的抗-JK a;152例意外抗体阴性标本,2种试剂分别正确检出149例和152例。两者的灵敏度为97.22%(70/72)vs 98.61%(71/72),特异度为98.02%(149/152)vs 100%(152/152),配对卡方检验结果表明,2种试剂盒的检测结果差异不具有统计学意义(P>0.05)。结论采用固相凝集法研发的试剂盒,适用于IgG型意外抗体的鉴定。Objective To develop a solid-phase red cell adherence(SPRCA)reagent kit for unexpected alloantibody identification.Methods A solid-phase carrier for bounding red blood cell(RBC)was prepared by coating microstrip with succinimide,then panel cell suspensions were dispensed to microwells to produce RBC monolayers,after that,1×PBS(pH7.40)was used to lysis the RBC monolayer to form stroma monolayer.Prior to the drying procedure,drying medium suspension was added to microwell to protect stroma monolayer.The solid-phase microplate was successfully produced after the stroma had been dried in refrigerator at 4℃.IgG anti-E and anti-Fy a with low titer were tested every two months with the reagent kit to observe its stability.Seventy-two plasma samples containing unexpected antibodies and 152 normal plasma samples were determined with SPRCA reagent kit and column agglutination technique(CAT).Performances including sensitivity and specificity of the two reagents were compared by paired chi-square test.Results RBC stroma monolayer could be attached well on microwell bottom coated with succinimide.Drying medium suspension could provide effective protection for stroma monolayer.Both IgG anti-E and anti-Fy a with low titer were determined well by the reagent kit during the storage period.Of the 72 positive samples,seventy 70 were identified correctly by SPRCA kit and 71 by CAT[Sensitivity:97.22%(70/72)vs 98.61%(71/72)].Of the 152 negative plasma samples,149 were correctly detected by SPRCA kit and 152 by CAT[Specificity:98.02%(149/152)vs 100%(152/152)].Paired chi-square test showed no significant differences between the two reagents(P>0.05).Conclusion A SPRCA reagent kit for IgG unexpected antibody identification had been developed successfully.

关 键 词：固相凝集法 意外抗体鉴定 试剂盒 

分 类 号：R457.1[医药卫生—治疗学]