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Application of microchip assay system for the measurement of C-reactive protein in human saliva

机译：芯片检测系统在人唾液中C反应蛋白测定中的应用

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In the last decade, saliva has been advocated as a non-invasive alternative to blood as a diagnostic fluid. However, use of saliva has been hindered by the inadequate sensitivity of current methods to detect the lower salivary concentrations of many constituents compared to serum. Furthermore, developments in the areas related to lab-on-a-chip systems for saliva-based point of care diagnostics are complicated by the high viscocity and heterogeneous properties associated with this diagnostic fluid. The biomarker C-reactive protein (CRP) is an acute phase reactant and a well-accepted indicator of inflammation. Numerous clinical studies have established elevated serum CRP as a strong, independent risk factor for the development of cardiovascular disease (CVD). CVD has also been associated with oral infections (i.e. periodontal diseases) and there is evidence that systemic CRP may be a link between the two. Clinical measurements of CRP in serum are currently performed with high sensitivity CRP (hsCRP) enzyme-linked immunosorbent assay (ELISA) tests that lack the sensitivity for the detection of this important biomarker in saliva. Because measurement of salivary CRP may represent a novel approach for diagnosing and monitoring chronic inflammatory disease, including CVD and periodontal diseases, the objective of this study was to apply an ultra-sensitive microchip assay system for the measurement of CRP in human saliva. Here, we describe this novel lab-on-a-chip system in its first application for the measurement of CRP in saliva and demonstrate its advantages over the traditional ELISA method. The increased sensitivity of the microchip system (10 pg ml~(-1) of CRP with 1000-fold dilution of saliva sample) is attributed to its inherent increased signal to noise ratio, resulting from the higher bead surface area available for antigen/antibody interactions and the high stringency washes associated with this approach. Finally, the microchip assay system was utilized in this study to provide direct experimental evidence that chronic periodontal disease may be associated with higher levels of salivary CRP.
机译：在过去的十年中，唾液被提倡作为血液作为诊断液的一种非侵入性替代方法。但是，唾液的使用由于当前方法检测许多成分的唾液浓度低于血清的敏感性不足而受到阻碍。此外，与基于唾液的即时诊断的芯片实验室系统有关的领域的开发由于与此诊断液相关的高粘度和异质性而变得复杂。生物标志物C反应蛋白（CRP）是急性期反应物，是炎症的公认指标。许多临床研究已将血清CRP升高确定为心血管疾病（CVD）发生的强大独立危险因素。 CVD也与口腔感染（即牙周疾病）有关，有证据表明全身CRP可能是两者之间的联系。目前，血清CRP的临床测量是使用高灵敏度CRP（hsCRP）酶联免疫吸附测定（ELISA）测试进行的，该测试缺乏检测唾液中这种重要生物标志物的灵敏度。由于唾液CRP的测量可能代表一种诊断和监测包括CVD和牙周疾病在内的慢性炎性疾病的新颖方法，因此本研究的目的是将超灵敏的微芯片测定系统应用于人唾液中CRP的测量。在这里，我们描述了这种新颖的单芯片实验室系统在其首次用于唾液中CRP测量的应用，并证明了其优于传统ELISA方法的优势。微芯片系统（10 pg ml〜（-1）的CRP和唾液样品稀释1000倍的稀释液）灵敏度的提高归因于其固有的信噪比提高，这是因为抗原/抗体可利用的磁珠表面积更高交互和与此方法相关的高严格性洗涤。最后，在本研究中使用微芯片测定系统提供直接的实验证据，证明慢性牙周疾病可能与唾液CRP水平升高有关。

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