Development and validation of an automated and ultrasensitive immunoturbidimetric assay for C-reactive protein.
Development and validation of an automated and ultrasensitive immunoturbidimetric assay for C-reactive protein.
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开发和验证 C 反应蛋白的自动化超灵敏免疫比浊测定法。
DOI：
10.1093/clinchem/46.11.1839
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发表时间：
2000
影响因子：
9.3
通讯作者：
Francisco Galisteo
中科院分区：
文献类型：
--
作者：
Luis Borque;L. Bellod;A. Rus;M.;Luisa Seco;Francisco Galisteo
C反应蛋白（CRP）是炎症急性期反应的标志物，在疾病发作后数小时内迅速增加。多年来，CRP测量已用于各种临床情况的管理，如细菌感染、组织缺血性坏死和活动性炎症（1）。最近的研究CRP浓度在传统的参考区间内提出了新的临床应用。CRP是心绞痛患者（稳定或不稳定）和明显健康受试者中冠心病的预后风险因素（2），并区分快速破坏性与缓慢进展性骨关节炎（3）。在新生儿中，CRP >1-2 mg/L可能与严重疾病相关，通常是细菌感染（4）。在大多数这些研究中，作者提出CRP的临界点为2-3 mg/L。最近的报告显示，健康对照组的中位值为0.58-1.72 mg/L，95百分位数范围为0.1-6.0 mg/L（5）（6）。然而，这些结果可能会受到CRP定量方法灵敏度的影响。
在许多市售的CRP检测方法中，最常用的是液相或颗粒增强比浊法或散射比浊法（7）。这种测定法适用于测量浓度>5-10 mg/L的蛋白质。测量CRP的新方法的检测下限是早期方法的十分之一（8）（9）。一些使用昂贵的单克隆抗体，而另一些则在反应混合物中使用更高的样品体积和抗体浓度，这会增加非特异性反应（10）。
我们描述了一种微粒试剂，我们使用在一个优化的浊度法与免疫纯化的多克隆抗体对CRP。它允许微粒试剂上的较低抗体覆盖率，并为血清CRP提供高度灵敏和稳健的微粒增强比浊免疫测定。此外，我们提出了一种新的顺序共价偶联IgG和牛血清白蛋白的程序。
C-Reactive protein (CRP) is a marker of acute phase response to inflammation that increases rapidly within hours of disease onset. CRP measurements have been used for many years in the management of a variety of clinical situations, such as bacterial infections, ischemic necrosis of tissue, and active inflammatory conditions (1). Recent studies of CRP concentrations within the conventional reference interval have suggested new clinical applications. CRP is a prognostic risk factor for coronary heart disease in both patients with angina (stable or not) and in apparently healthy subjects (2), and distinguishes rapidly destructive vs slowly progressive osteoarthritis (3). In neonates, CRP >1–2 mg/L could be associated with serious disease, usually bacterial infection (4). In most of these studies, the authors have proposed a cutoff point for CRP of 2–3 mg/L. Recent reports show median values in healthy controls of 0.58–1.72 mg/L, and 95 percentile ranges of ∼0.1–6.0 mg/L (5)(6). Nevertheless, these results can be influenced by the sensitivity of the methodology used for CRP quantification.
Among the many commercially available assays for CRP, the most common use fluid-phase or particle-enhanced turbidimetric or nephelometric procedures (7). Such assays are suitable for the measurement of proteins at concentrations >5–10 mg/L. New methods of measuring CRP have lower limits of detection one-tenth those of earlier methods (8)(9). Some use expensive monoclonal antibodies, and others use higher sample volumes and antibody concentrations in the reaction mixtures, which can increase nonspecific reactions (10).
We describe a microparticle reagent that we use in an optimized turbidimetric procedure with immunopurified polyclonal antibodies against CRP. It allows lower antibody coverage on the microparticle reagent and provides highly sensitive and robust particle-enhanced turbidimetric immunoassay for serum CRP. In addition, we present a new procedure of sequential covalent coupling of IgG and bovine serum albumin …
影响因子：
9.3
作者：
E. Macy;T. Hayes;R. Tracy
通讯作者：
E. Macy;T. Hayes;R. Tracy