Develop a solid-phase red cell adherence reagent kit for IgG unexpected alloantibody identification
Objective To develop a solid-phase red cell adherence(SPRC A)reagent kit for unexpected alloantibody i-dentification.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-Fya 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 sensi-tivity 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-Fya with low titer were determined well by the reagent kit during the storage peri-od.Of the 72 positive samples,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[Spe-cificity: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 suc-cessfully.
万方数据
维普
