A high sensitivity assay for the inflammatory marker C-Reactive protein employing acoustic biosensing.
A high sensitivity assay for the inflammatory marker C-Reactive protein employing acoustic biosensing.
复制标题
DOI：
10.1186/1477-3155-6-5
复制
发表时间：
2008-04-29
影响因子：
10.2
通讯作者：
Cooper MA
中科院分区：
文献类型：
--
作者：
McBride JD;Cooper MA
C-反应蛋白（CRP）是一种急性期反应物，通常用作生物标志物，以评估感染或炎症过程，如自身免疫性疾病。CRP也被证明是未来心血管疾病风险的预测标志物。已经开发了一种用于检测C-反应蛋白的免疫测定的新方法，其使用共振声学分析™（RAP™），具有与高灵敏度CRP ELISA（hsCRP）相当的灵敏度，但具有相当的时间效率（12分钟的周转时间到结果）。在一种方法中，通过RAP™生物传感器的两个传感器通道注射CRP的标准溶液（0至231 ng/mL）或稀释的掺加马血清样品。一个包含具有针对CRP的绵羊抗体的表面，另一个包含纯化的绵羊IgG的对照表面。在5分钟注射结束时，共振频率的初始变化率与CRP浓度成比例。抗CRP结合的第二个夹心步骤的初始速率也与样本CRP浓度成比例，并提供了一种更灵敏的CRP定量方法。直接试验和均质夹心试验的检测下限均为20 ng/mL，而直接夹心试验的检测下限为3 ng/mL。在快速临床测定的一个步骤中，加标人CRP的稀释马血通过一个传感器通道，而临界心血管风险水平的参比标准品溶液通过另一个通道。因此获得了指示样本CRP状态的半定量比率。总体而言，本研究揭示了在正常和病理条件下可能预期的血清中的CRP浓度可以在具有RAP™检测技术的时间有效的无标记免疫测定中检测到，其中所确定的CRP浓度与使用市售高灵敏度ELISA所确定的CRP浓度非常一致。
C-Reactive Protein (CRP) is an acute phase reactant routinely used as a biomarker to assess either infection or inflammatory processes such as autoimmune diseases. CRP also has demonstrated utility as a predictive marker of future risk of cardiovascular disease. A new method of immunoassay for the detection of C-Reactive Protein has been developed using Resonant Acoustic Profiling™ (RAP™) with comparable sensitivity to a high sensitivity CRP ELISA (hsCRP) but with considerable time efficiency (12 minutes turnaround time to result). In one method, standard solutions of CRP (0 to 231 ng/mL) or diluted spiked horse serum sample are injected through two sensor channels of a RAP™ biosensor. One contains a surface with sheep antibody to CRP, the other a control surface containing purified Sheep IgG. At the end of a 5-minute injection the initial rate of change in resonant frequency was proportional to CRP concentration. The initial rates of a second sandwich step of anti-CRP binding were also proportional to the sample CRP concentration and provided a more sensitive method for quantification of CRP. The lower limit of detection for the direct assay and the homogenous sandwich assay were both 20 ng/mL whereas for the direct sandwich assay the lower limit was 3 ng/mL. In a step towards a rapid clinical assay, diluted horse blood spiked with human CRP was passed over one sensor channel whilst a reference standard solution at the borderline cardiovascular risk level was passed over the other. A semi-quantities ratio was thus obtained indicative of sample CRP status. Overall, the present study revealed that CRP concentrations in serum that might be expected in both normal and pathological conditions can be detected in a time-efficient, label-free immunoassay with RAP™ detection technology with determined CRP concentrations in close agreement with those determined using a commercially available high sensitivity ELISA.