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血清脂蛋白(a)速率散射比浊法测定

Rate nephelometric assay of serum lipoprotein(a).

作者信息

Gillery P, Arthuis P, Cuperlier C, Circaud R

机构信息

Laboratory of Biochemistry, Centre Hospitalier Universitaire de Reims, Hôpital Robert Debre, France.

出版信息

Clin Chem. 1993 Mar;39(3):503-8.

PMID:8448865
Abstract

This nephelometric assay of serum lipoprotein(a) [Lp(a)] is characterized by the use of a specific antibody to generate a high rate of light-scatter formation and the elimination of nonspecific reactions from serum samples by diluting samples in phosphate-buffered saline containing polymer enhancer polyethylene glycol (PEG), 40 g/L, and detergent before the assay. We reacted 100 microL of sixfold-diluted serum in 500 microL of buffer containing PEG with 42 microL of pure polyclonal rabbit antiserum (Dakopatts) directed against human Lp(a) and monitored the reaction by rate nephelometry with the Array Protein System nephelometer (Beckman). The standard curve for the reaction was linear in the Lp(a) range 10-1280 mg/L; antigen excess occurred between 1300 and 1400 mg/L. Calibration was performed with serial dilutions of a standard serum. Precision studies showed within-run and between-run CVs of < 2.1% and 6.9%, respectively. The nephelometric results (y) for 100 serum samples were highly correlated with those obtained by radial immunodiffusion (x) calibrated with the same materials: y = 1.07 (+/- 0.03) x - 16.2 (+/- 8.1) mg/L (r = 0.974, P < 0.001). Storing serum for 3 weeks at 4 degrees C or 3 months at -80 degrees C did not affect the results.

摘要

这种血清脂蛋白(a)[Lp(a)]的散射比浊测定法的特点是,使用特异性抗体产生高散射光形成速率,并在测定前通过在含有40 g/L聚合物增强剂聚乙二醇(PEG)和去污剂的磷酸盐缓冲盐水中稀释血清样本,消除血清样本中的非特异性反应。我们将100 μL六倍稀释的血清与500 μL含有PEG的缓冲液混合,再加入42 μL针对人Lp(a)的纯多克隆兔抗血清(Dakopatts),并使用Array Protein System散射比浊仪(贝克曼)通过速率散射比浊法监测反应。该反应的标准曲线在Lp(a)浓度范围10 - 1280 mg/L内呈线性;在1300至1400 mg/L之间出现抗原过量。用标准血清的系列稀释液进行校准。精密度研究表明批内和批间变异系数分别<2.1%和6.9%。100份血清样本的散射比浊结果(y)与用相同材料校准的放射免疫扩散法得到的结果(x)高度相关:y = 1.07(±0.03)x - 16.2(±8.1)mg/L(r = 0.974,P < 0.001)。血清在4℃储存3周或在-80℃储存3个月不影响结果。

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