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[放射性抗球蛋白(库姆斯氏)试验]

[The radioactive antiglobulin (Coombs') test].

作者信息

Spath P, Yam P, Petz L D

出版信息

Acta Med Austriaca. 1979;6(5):204-6.

PMID:555218
Abstract

Weak red cell sensitization by hemolyzing antibodies may not be detected by the standard antiglobulin test. For the diagnosis of certain immune hemolytic anemias more sensitive methods are needed. Anti-IgG was purified as F(ab)2 by pepsin digestion and labelled with 125 I using the chloramine T method. 15 X 10(7) cells were washed and suspended in 0.1 ml of 6% albumin in saline. After incubation with 0.1 ml of 125 I-anti-IgG F(ab')2 for 30 minutes, the cells were washed and the uptake of radioactivity was counted using a gamma-counter. Standard cells were coated with a known concentration of an IgG anti-D preparation in the range of 50--1000 molecules per red cell. The sensitivity for the detection of red cell sensitization was at least 50 molecules per red cell. Because of varying binding ratios between the labelled anti-IgG reagent and different red cell antibodies the determination of the exact number of IgG molecules on the red cells is difficult. However, this very sensitive and reproducible method shows practical advantages in comparison with a complement fixation antibody consumption method with which corresponding results were obtained. Beside the evaluation of antiglobulin test negative autoimmune hemolytic anemias, this method may be applied for the determination of the specificity of weak red cell alloantibodies not detectable by standard blood bank procedures.

摘要

溶血抗体引起的弱红细胞致敏可能无法通过标准抗球蛋白试验检测到。对于某些免疫性溶血性贫血的诊断,需要更敏感的方法。通过胃蛋白酶消化将抗IgG纯化至F(ab)2,并使用氯胺T法用125I进行标记。将15×10(7)个细胞洗涤后,悬浮于0.1ml含6%白蛋白的生理盐水中。与0.1ml 125I-抗IgG F(ab')2孵育30分钟后,洗涤细胞并使用γ计数器计数放射性摄取量。标准细胞用已知浓度的IgG抗-D制剂包被,浓度范围为每个红细胞50-1000个分子。检测红细胞致敏的灵敏度至少为每个红细胞50个分子。由于标记的抗IgG试剂与不同红细胞抗体之间的结合比例不同,难以确定红细胞上IgG分子的确切数量。然而,与获得相应结果的补体结合抗体消耗法相比,这种非常敏感且可重复的方法具有实际优势。除了评估抗球蛋白试验阴性的自身免疫性溶血性贫血外,该方法还可用于确定标准血库程序无法检测到的弱红细胞同种抗体的特异性。

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