Development of rapid analysis device for periodontal disease activity

牙周病活动度快速分析装置的研制

基本信息

批准号：
18592262
负责人：
KATAOKA Masatoshi
金额：
$ 2.49万
依托单位：
National Institute of Advanced Industrial Science and Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

Several biomarkers including procollagen type I C-terminal propeptide(PICP), have been reported for objective evaluation of the state of periodontal disease. Although these markers are usually examined by conventional ELISA method with a microplate reader, it is time-and sample-consuming, and it is not adaptable to automated analyzers. Convenient, sensitive, and accurate methods for the measurement of the biomarkers are thus required. Bio-nano technology has received considerable interest in analytical chemistry due to the intrinsic characteristics of high speed, high throughput, easy operation, low consumption of samples and reagents, miniaturization, and automation. To develop the analysis system of biomarkers in periodontal disease in dental clinic, we first developed the quantitative analysis system based on the antigen-antibody reaction in microchannel on microchip. After the adsorption of 1st anti-PICP antibody on the surface of microchannel (300 μm width, 100 μm depth) on cyclic olefin copolymers (COC) microchip, sample and peroxidase-conjugated 2nd anti-PICP antibody mixture was introduced into a microchannel for 30 min, followed by washing and peroxidase activity was measured for the determination of PICP level. A double-logarithm plot analysis revealed a linear relationship between PICP concentration and peroxidase activity in the range of 50 to 600 ng/ml of PICP (r^2=0.9745). PICP concentrations in humoris of four subjects determined by the present methods were compared with the results obtained by conventional ELISA method. Good correlations were observed for each method on simple linear regression analysis (P<0.05). We demonstrated the possible application of microchip for determination of PICP levels with high sensitivity and accuracy, and the potential for the dental clinical diagnosis.

已经报道了几种生物标志物在内，包括I型C末端前肽（PICP），用于客观评估牙周疾病的状态。尽管这些标记通常通过传统的ELISA方法和微型读取器进行检查，但它是耗时的，并且不适合自动分析仪。因此，需要方便，敏感和准确的生物标志物测量方法。由于高速，高吞吐量，易于操作，样品和试剂的消耗率低，小型化和自动化的内在特征，因此对分析化学的兴趣已引起了人们的兴趣。为了开发牙科诊所牙周疾病中生物标志物的分析系统，我们首先基于微芯片上微通道中的抗原抗体反应开发了定量分析系统。在对微通道（300μm宽度，100μM深度）表面吸附后，在环状烯烃共聚物（COC）微芯片上，样品和过氧化物酶 - 偶联的第二抗PICP抗体混合物中，在30分钟中，将其引入了30分钟的启用，在环环烯烃共聚物（COC）微芯片（COC）微芯片（COC）微芯片（COC）微芯片（COC）微芯片（COC）微芯片（COC）微芯片（COC）微芯片（COC）微芯片（COC）微芯片（COC）微芯片上，以; PICP级别。双层图分析揭示了PICP浓度和过氧化物酶活性之间的线性关系，范围为50至600 ng/ml的PICP（R^2 = 0.9745）。将通过本方法确定的四个受试者的Humoris中的PICP浓度与传统ELISA方法获得的结果进行了比较。对于简单的线性回归分析，每种方法都观察到良好的相关性（P <0.05）。我们证明了微芯片可能在确定具有高灵敏度和准确性的PICP水平以及牙科临床诊断的潜力。