改良DTT法用于减少达雷妥尤单抗对血清学检测造成的干扰

Application of Modified DTT Methods to Reduce the Interference of Daratumumab on Serological Detection

目的:改良美国输血协会《AABB技术手册》中经典的二硫苏糖醇(DTT)处理红细胞法,探讨其在使用达雷妥尤单抗(daratumumab)治疗的患者输血前检查中的应用价值。方法:从PBS、DTT浓度、供者红细胞浓度(悬浮/压积)及样本处理时间方面,对经典的0.2 mol/L AABB DTT法进行改良,将改良DTT法和AABB经典DTT方法应用于12例达雷妥尤单抗治疗的多发性骨髓瘤患者的配血试验,并用已知抗体性质的抗血清和抗体试剂评估改良DTT法处理谱红细胞后对其他不规则抗体检出的影响。结果:建立了2种改良DTT法(方法一:将DTT浓度改为0.01 mol/L;方法二:将DTT浓度改为0.02 mol/L,并将压积红细胞改为3%悬浮红细胞)。改良DTT法处理红细胞最佳处理时间为35 min,不仅能够消除达雷妥尤单抗造成的全凝集现象,且不影响抗-E、抗-Jk^(a)、抗-M等抗体的检出,对抗-K抗体的测定效价仅有略微下降。结论:改良DTT法安全有效,能够在消除达雷妥尤单抗干扰的同时保留Kell血型系统活性,可以替代目前AABB经典的DTT配血方法。

Objective:To modified the classic dithiothreitol(DTT)method for treating red blood cells(RBCs)in Technical Manual of American Association of Blood Banks(AABB)and evaluate its application value in pre-transfusion examination of patients treated with daratumumab.Methods:The classic 0.2 mol/L DTT method was improved in terms of PBS,DTT concentration,donor RBCs concentration(suspended/packed)and sample processing time.The modified DTT methods and AABB classic DTT method were applied to the blood matching tests of 12 multiple myeloma patients treated with daratumumab.The effect of treating panel RBCs with modified DTT methods on the detection of other irregular antibodies was evaluated by using antiserum and antibody reagents with known antibody properties.Results:Two modified DTT methods were established(method 1:changed the concentration of DTT to 0.01 mol/L;method 2:changed the concentration of DTT to 0.02 mol/L and replaced the packed RBCs with 3%RBCs suspension).The optimal treatment time was 35 min for the modified DTT methods.At this time,the pan-agglutination caused by daratumumab was eliminated,but the detection of antibodies such as anti-E,anti-Jka,anti-M were not affected,and the titer of anti-K antibodies was only slightly decreased.Conclusion:The modified DTT methods were effective,which can eliminate the interference of daratumumab while retaining the activity of the Kell blood group system,and can replace the current classic DTT method in AABB Technical Manual.