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使用可磁化纤维素颗粒的C反应蛋白固相免疫放射分析

Solid-phase immunoradiometric assay for C-reactive protein using magnetisable cellulose particles.

作者信息

De Beer F C, Pepys M B

出版信息

J Immunol Methods. 1982;50(3):299-308. doi: 10.1016/0022-1759(82)90168-5.

Abstract

An immunoradiometric assay (IRMA) for C-reactive protein (CRP) was developed using magnetisable cellulose particles as the solid-phase support for anti-CRP antibodies. 125I-labeled immunopurified anti-CRP antibody was used to quantitate the amount of CRP taken up by the solid phase. Unbound label was easily and rapidly removed by decantation after sedimenting the particles on a magnet. The assay could detect 1 microgram CRP/l and had a range of up to 10 mg/l with the portion of the standard curve between 10 micrograms/l and 2-3 mg/l being linear. Fifty samples per hour could be processed manually from serum to CRP result with an intra-assay CV of 5.2% and an inter-assay CV of 10.0%, based on 5 replicates of 5 samples with CRP levels between 2 mg/l and 180 mg/l run in 5 separate assays. Fifty clinical samples were assayed in parallel with a standard electroimmunoassay and yielded a linear correlation coefficient (r) of 0.975 and a slope of 0.988. With its single, brief incubation step including all reagents and its simple phase separation procedure the present method may be the assay of choice when precise measurement of CRP concentrations is required rapidly.

摘要

一种用于检测C反应蛋白(CRP)的免疫放射分析(IRMA)方法被开发出来,该方法使用可磁化的纤维素颗粒作为抗CRP抗体的固相支持物。用125I标记的免疫纯化抗CRP抗体来定量固相所摄取的CRP量。在将颗粒用磁铁沉淀后,通过倾析可轻松快速地去除未结合的标记物。该检测方法能够检测到1微克CRP/升,线性范围高达10毫克/升,标准曲线在10微克/升至2 - 3毫克/升之间呈线性。基于在5次独立检测中对5个CRP水平在2毫克/升至180毫克/升之间的样本进行5次重复检测,每小时可手动处理50个样本,从血清得出CRP结果,批内变异系数(CV)为5.2%,批间变异系数为10.0%。50份临床样本与标准免疫电泳分析同时进行检测,得到的线性相关系数(r)为0.975,斜率为0.988。由于本方法具有包括所有试剂的单一简短孵育步骤以及简单的相分离程序,在需要快速精确测量CRP浓度时,它可能是首选的检测方法。

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