Integrated PCR Diagnostic and Analysis Instrument

描述（申请人摘要）：聚合酶链反应的力量 (PCR)作为一种分析工具已经得到了广泛的测试。近年来，许多临床 诊断程序，特别是用于筛选几种类型的癌症， 广泛的细菌、病毒和寄生虫病， 亲子鉴定、法医分析和组织分型中的鉴定 开始使用PCR技术。尽管14年前发明了PCR， 大多数研究人员和诊断实验室一步一步地进行PCR分析， 每一步都有手动传输和专用的单功能仪器。 此外，大量的人工处理和物理传热 在热循环步骤中的限制，可以有助于1/2到两天长 PCR检测。尽管有一些自动化工具可用于减少 每一步都是手工操作，目前没有成本效益高的仪器 以灵活的方式完全自动化PCR方案。此相反 许多完全自动化的非PCR诊断仪器， 现代临床实验室我们建议将最近的几项进展应用于小 PCR的样品处理、快速热循环和一般样品处理 协议，以加快分析速度，降低成本和体力劳动。一 技术人员只需将一个DNA模板板和一个 所需的PCR引物，然后启动仪器。该仪器将精确地 吸取所需体积的每个样品，然后进行热循环， PCR后清理、检测和结果跟踪步骤， 自动化的方式，提供全自动化。该文书将减少整个 测试成本（试剂、一次性用品和劳动力）增加20倍或更多。 考虑到PCR技术的广泛应用，我们提出将该仪器 柔性的（例如，不同类型的检测协议， 仪器硬件）。新工具的基础是执行所有 功能：混合、热循环、清洁、检测和灭菌 在同一组纳米移液器反应容器中。在第一阶段， 目标是两个基于PCR的方案：一个用于DNA诊断，一个用于SNP 分析.拟议的仪器的主要特点，即快速， 小体积PCR、荧光检测和反应室清洁将 在第一阶段项目中开发和演示。第二阶段将需要 开发完整的原型仪器， 使用选定的方案对样品进行低容量处理。 拟定商业应用： 这项工作将导致先进性能的商业PCR分析仪器。 这将使研究人员能够使用高通量技术更快地做出重要发现。 筛选试验，并成为一个有效的和成本效益的遗传和 传染病诊断

DESCRIPTION (Applicant's abstract): The power of polymerase chain reaction (PCR) as an analytical tool has been widely tested. Recently, many clinical diagnostic procedures used especially for screening several types of cancer, a wide range of bacterial, viral and parasitic diseases, individual identification in paternity testing, forensic analysis, and tissue typing have started employing PCR technology. Despite the invention of PCR 14 years ago, most researchers and diagnostic labs conduct PCR analyses one step at a time, with manual transfers and dedicated, single-function instruments at each step. Furthermore, both extensive manual handling and the physical heat-transfer constraints in the thermal cycling step, can contribute to 1/2 to two day long PCR tests. Although there are some automated tools available to reduce the manual work at each step, there are currently no cost effective instruments that completely automate the PCR protocol in a flexible manner. This contrasts with many of the completely automated non-PCR-based diagnostic instruments in the modern clinical labs. We propose to apply several recent advances in small sample handling, fast thermal cycling and general sample processing to the PCR protocol in order to speed analysis and reduce costs and manual labor. A technician would simply load a plate of DNA templates and a plate of the required PCR primers, then start the instrument. The instrument would precisely aspirate the required volume of each sample, then perform the thermal cycling, post-PCR cleanup, detection and results-tracking steps in a completely automated fashion, offering full automation. The instrument would reduce entire test costs (reagents, disposables and labor) by a factor of 20 or more. Considering the wide applications of PCR, we propose to make the instrument flexible (e.g., different types of detection protocols with few changes to the instrument hardware). The basis of the new instrument is to perform all functions: mixing, thermal cycling, clean up, detection and sterilization within the same set of nano-pipetter reaction vessels. In the Phase I, the target is on two PCR-based protocols: one for DNA diagnostics and one for SNP analysis. The key features of the proposed instrument, namely fast, small-volume PCR, fluorescent detection and reaction chamber cleaning will be developed and demonstrated within the Phase I project. Phase II will entail the development of a complete prototype instrument and demonstration of high-speed, low-volume processing of samples using selected protocols. PROPOSED COMMERCIAL APPLICATION: This work will result in advanced-performance commercial PCR analysis instruments. These will allow researchers to more rapidly make important discoveries using high-throughput screening assays and be the basis for an efficient and cost effective system for genetic and infectious disease diagnosis.