Nelson M, Zarkos K, Popp H, Gibson J
Institute of Haematology, Royal Prince Alfred Hospital, Sydney, N.S. W., Australia.
Vox Sang. 1998;75(3):234-41.
The Kleihauer slide test is in general use to screen obstetric patients for possible fetomaternal haemorrhage. Since 1993, Rh(D)-negative patients have been tested in our laboratory by a flow-cytometric method detecting Rh(D)-positive fetal cells, a method which offers improved sensitivity and accuracy. We report another flow-cytometric method of broader application which quantitates cells according to haemoglobin F (HbF) content.
The red cells are fixed with glutaraldehyde and permeabilized by exposure to Triton X-100. A polyclonal sheep antibody to HbF is incubated with the cells followed by a fluorescein-labelled anti-sheep antibody.
Quantitation of the percentage of fetal cells following a FMH can be achieved irrespective of the blood groups of either mother or infant, and the presence of maternal F cells need not interfere since the intensity of staining is usually less than that of fetal cells. Two of 19 transfusion-dependent patients with beta-thalassaemia have been found to have red cells indistinguishable from fetal cells on the basis of HbF content, but these patients also have been found to give positive results by the Kleihauer test.
The flow-cytometric method may serve to replace the traditional Kleihauer test since it appears to offer improved accuracy and objectivity.
克莱豪尔玻片试验通常用于筛查产科患者是否可能发生胎儿-母体出血。自1993年以来，我院实验室对Rh(D)阴性患者采用流式细胞术检测Rh(D)阳性胎儿细胞，该方法具有更高的灵敏度和准确性。我们报告了另一种应用更广泛的流式细胞术方法，该方法根据血红蛋白F（HbF）含量对细胞进行定量分析。
红细胞用戊二醛固定，并用Triton X-100通透处理。将抗HbF的多克隆羊抗体与细胞孵育，然后加入荧光素标记的抗羊抗体。
无论母亲或婴儿的血型如何，均可对胎儿-母体出血后胎儿细胞的百分比进行定量分析，且母体F细胞的存在不会产生干扰，因为其染色强度通常低于胎儿细胞。在19例依赖输血的β地中海贫血患者中，有2例患者的红细胞根据HbF含量与胎儿细胞难以区分，但这些患者的克莱豪尔试验结果也呈阳性。
流式细胞术方法似乎具有更高的准确性和客观性，可用于取代传统的克莱豪尔试验。
