放散试验室内质控方法的初步探讨

Preliminary discussion on internal quality control for elution test

目的:建立一种简单有效、可在一般实验室使用的放散试验质控方法。方法:采用供者直接抗人球蛋白试验阴性、自身对照阴性、无免疫史的O型RhD阳性压积红细胞制备放散试验的阴性对照。采用不同稀释倍数的单克隆IgG型抗-D致敏O型RhD阳性红细胞制备强阳性、弱阳性2个梯度的放散试验阳性对照。采用正交试验原理摸索制备阳性对照的最佳制备条件。评价该质控方法对酸、热放散2种常用方法的适用性。结果:筛选到未致敏的O型RhD阳性压积红细胞作为放散试验的阴性对照。强阳性对照的最佳制备条件:200μL未致敏的O型RhD阳性压积红细胞与25μL的单克隆IgG型抗-D原液在37℃孵育1 min。弱阳性对照的最佳制备条件:200μL未致敏的O型RhD阳性压积红细胞与等体积的256倍稀释的单克隆IgG型抗-D在37℃孵育1 min。该质控方法只适用于酸放散,不适用于热放散。结论:成功建立了适用于酸放散的室内质控方法。

Objective:To establish a simple and effective internal control strategy for elution test in general laboratories. Method:The negative control of elution test was prepared by O/RhD-positive packed red cells which were direct antiglobulin test negative,self-control negative and from donors without immune history.Two levels of positive controls were processed by different concentrations of IgG anti-D reagent.The optimal conditions for the preparation of positive controls were determined through orthogonal tests.And the internal control strategy was evaluated for both acid elution and thermal elution test. Result:The negative control of elution test was successfully obtained with non-sensitized O/RhD-positive packed red cells.The optimal conditions of strong positive control,200 μL non-sensitized O/RhD-positive packed red cells were well incubated with 25 μL IgG anti-D reagent at 37℃ for 1 min.The optimal conditions of weakly positive control,200 μL non-sensitized O/RhD-positive packed red cells were well incubated with 200 μL diluted IgG anti-D reagent(dilution at a ratio of 1:256)at 37℃ for 1 min.Our quality control method was applicable only to acid elution test,but not thermal elution test.Conclusion:An internal quality control strategy is successfully established for acid elution test.