超离心血清激光浊度法简化免疫复合物筛选。

Diagnostic immunology Pub Date : 1984-01-01
J R Cohn, C E Buckley, C D Connell
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引用次数: 0

摘要

14名健康志愿者在禁食一夜后和饭后再次抽取血清样本。每个样品被分成两个等分。一个等分液以20000 rpm离心20分钟,通过穿刺管的侧面从管的中间取出一部分。然后将所有样品加入单独含有0.14 M NaCl或0.15 M KCl中含有1,2,3或4%聚乙二醇的比色皿中。测量了每个管的光散射。与未离心的对照组相比,离心后的样品散射的光少了58%。用离心样品得到的值的范围也较小,这反映在每个聚乙二醇浓度下得到的平均值的标准偏差上。即使样品处理方式不同,重复样品的变异系数也在0 ~ 16.6%(平均5.8%)之间。与基线值相比,-70℃的储存显著增加了光散射。从44名患有与免疫复合物相关疾病的患者中抽取血清样本。浊度法选择性地识别出一些异常的患者。我们得出的结论是,在不沉淀出免疫复合物的条件下,制备性超离心消除了血清样品的大部分背景光散射,包括脂质散射。因为它测量的是免疫复合物的物理特性,所以这种检测方法可以提供基于生物受体的检测方法无法提供的信息。
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Simplified screening for immune complexes by laser nephelometry of ultracentrifuged serum.

Serum samples were drawn from 14 healthy volunteers after an overnight fast and again after a meal. Each sample was divided into two aliquots. One aliquot was centrifuged at 20,000 rpm for 20 min, and a portion from the middle of the tube was removed by puncturing the tube's side. All samples were then added to cuvettes containing 0.14 M NaCl alone or 1, 2, 3, or 4% polyethylene glycol in 0.15 M KCl. The light scattering of each tube was measured. The centrifuged samples scattered 58% less light than the uncentrifuged controls. The range of values obtained with the centrifuged samples was also smaller as reflected in the standard deviation of the means obtained at each polyethylene glycol concentration. The coefficient of variation for replicate samples ranged from 0 to 16.6% (mean 5.8%), even when samples were processed differently. Storage at -70 degrees C significantly increased light scattering compared to baseline values. Serum samples were drawn from 44 patients with diseases that have been associated with immune complexes. Nephelometry selectively identified some groups of patients as abnormal. We conclude that preparative ultracentrifugation removes much of the background light scattering of serum samples, including that due to lipid, under conditions that do not sediment out immune complexes. Because it measures a physical property of immune complexes, this assay may provide information that is not available from biological receptor-based assays.

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