牙周病原体的对流型在线PCR-毛细管电泳检测方法

项目名称： 牙周病原体的对流型在线PCR-毛细管电泳检测方法

项目编号： No.21205078

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 分析化学

项目作者： 李振庆

作者单位： 上海理工大学

项目金额： 25万元

中文摘要： 牙周病是由龈下菌斑引起的一种慢性疾病，是危害牙齿健康的第二大口腔疾病，及时发现牙周病原体并对其加以控制，对牙齿健康具有重要的作用。本研究拟在实验室自制毛细管电泳(CE)系统的基础上，建立可实现DNA扩增的、多通道、Rayleigh-Bérd(R-B)自然对流的PCR-CE联用检测新方法。为此，本项目拟：(1) 通过数值模拟，设计与CE系统匹配的R-B对流腔体；(2)研究腔体几何尺寸、内部结构及边界条件对自然对流稳定性的影响以及腔体内不同位置处液体温度随时间的变化，建立可反映PCR腔体内传热过程的数学模型，为实现稳定的R-B对流和有效DNA扩增提供理论基础；(3) 利用所建系统完成牙周致病菌种Pg, Tf, Td的检测，揭示其含量与牙周病之间的内在联系。该方法的主要优点是系统结构简单，取样体积极小，多通道，对于细菌菌种鉴定及实现疾病床旁诊断POCT具有重要意义。

中文关键词： 牙周病原菌；聚合酶链式反应；毛细管电泳；自然对流；生物芯片

英文摘要： Periodontal disease, which is a chronic disease caused by subgingival plaque, is the second major oral disease endangering the dental health. Thus timely detection of periodontal pathogens and its control are of vital importance on the dental health. On the basis of lab-built capillary electrophoresis (CE) system, this project aims to develop the multi-channel, online polymerase chain reaction (PCR)-CE system with Rayleigh-Bérd (R-B)convection method. So, our plans for this project are as the followings: (1) Design the R-B convection column chamber which matches well with the CE system by numerical simulation. (2) We prepare to investigate the effect of the chamber dimensions, chamber configuration, and boundary conditions on the PCR process. Meanwhile, we will reseach the temperature changing with time at different sites of the chamber. Finally, we will build the mathematical modeling for the thermal transfer process, and thus to provide a theoreical basis for the stable PCR process with convection RLB. (3) We will attepmt to detect the periodontal pathogens, such as Porphyomonas gingivalis (Pg), Tannerella forsythia (Tf), and Treponema denticola (Td) on the PCR-CE system in this programme. We hope we can explore the quantitative relationship between periodontal pathogens in saliva of periodontal pocket and p

英文关键词： Periodontal pathogens；Polymerase chain reaction；Capillary electrophoresis；Natural convection；Biochip