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.

据报道，包括 I 型前胶原 C 末端前肽 (PICP) 在内的多种生物标志物可用于客观评估牙周疾病的状态，尽管这些标志物通常通过酶标仪的传统 ELISA 方法进行检查，但这种方法既耗时又耗时。生物标记物的测量方法不适合自动化分析仪，因此，生物纳米技术因其高速、高通量的内在特性而受到了分析化学的广泛关注。为了开发牙科诊所牙周病生物标志物分析系统，我们首先开发了基于微芯片上微通道抗原抗体反应的定量分析系统。将第一抗 PICP 抗体吸附在环烯烃共聚物 (COC) 微芯片上的微通道（300 μm 宽，100 μm 深）表面后，样品和将过氧化物酶缀合的第二抗 PICP 抗体混合物引入微通道中 30 分钟，然后洗涤并测量过氧化物酶活性以确定 PICP 水平，双对数图分析显示 PICP 浓度与过氧化物酶活性之间存在线性关系。 4 名受试者体液中 PICP 浓度范围为 50 至 600 ng/ml（r^2=0.9745）。将本方法与常规 ELISA 方法获得的结果进行比较，在简单线性回归分析中观察到每种方法具有良好的相关性（P<0.05），我们证明了微芯片用于高灵敏度和准确度测定 PICP 水平的可能性。牙科临床诊断的潜力。