Integrated molecular diagnostic system for the point-of-care

用于现场护理的综合分子诊断系统

基本信息

批准号：
8339922
负责人：
Huimin Kong
金额：
$ 24.09万
依托单位：
BIOHELIX CORPORATION
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-30 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8339922
关键词：
Bedside Testings Biological Assay Boston Buffers Centers for Disease Control and Prevention (U.S.)Chemistry Chlamydia trachomatis Client Clinic Clinical Cytolysis DNA DNA amplification Detection Device Designs Devices Diagnostic Engineering Eye Hand Health Personnel Housing Laboratories Lateral Learning Liquid substance Literature Manufacturer Name Marketing Microfluidic Microchips Molecular Movement National Institute of Allergy and Infectious Disease Neisseria gonorrhoeae Nucleic Acids Patients Phase Point-of-Care Systems Polymerase Chain Reaction Process Reaction Reader Reagent Research Risk Sampling Sexually Transmitted Diseases Small Business Technology Transfer Research Solid Solutions Staging Surgical Flaps System Technical Expertise Technology Temperature Test Result Testing Tube Universities Urine Validation Vent Visual Work base commercialization cost design experience fluorescence microscope helicase high throughput screening inhibitor/antagonist instrument internal control micro-total analysis system pathogen point of care prototype

项目摘要

DESCRIPTION (provided by applicant): This STTR-AT-NIAID seeks to develop an integrated nucleic acid system based on research done by Catherine Klapperich's laboratory at Boston University. The BU lab-on-a-chip includes a micro solid phase extraction (<SPE) column, flap valves and hydrophobic vents to gate fluid movements through micro channels, and multiple reaction chambers for experimental replicates and control reactions. Dr. Klapperich's laboratory has successfully performed nucleic acid amplifications in the chip using BioHelix's isothermal amplification process. Biohelix's proprietary technology is called helicase-dependent amplification (HDA). It uses DNA helicases to separate DNA strands during exponential amplification at a constant temperature of 65:C. Like the polymerase chain reaction (PCR), HDA assays use a competitive internal control (i.e., a template DNA of known concentration spiked into the raw sample) that can be amplified by the same primers as the analyte, but detected separately; hence allowing us to detect amplification inhibitors in direct clinical samples. The objectives of Phase I are to: 1) modify the BU microfluidic device design to incorporate a lateral flow strip as a means of detecting amplification products using the naked eye; and 2) demonstrate the feasibility of performing integrated tests using the device from aim 1 using CT and NG spiked at 104 in 1 mL urine to establish proof of concept for integrated assays performed in a modified BU chip that includes a lateral flow strip. Our specific aims for Phase II will be to develop a lateral flow reader sub-system, integrate it with the BU instrument to build a pre-commercial looks-like / works-like prototypes, simplify chip design and fabrication to allow for pilot scale manufacturing, and implement a pilot scale manufacturing for the disposable at BioHelix / Quidel. At this stage Qiagen is the most likely manufacturer for the reader. At the conclusion of Phase II, we should be ready for clinical validation of the new assay system.

描述（由申请人提供）：该 STTR-AT-NIAID 旨在基于波士顿大学 Catherine Klapperich 实验室所做的研究开发集成核酸系统。 BU 芯片实验室包括微固相萃取 (<SPE) 柱、瓣阀和疏水通风口（用于控制流体通过微通道的运动），以及用于实验重复和控制反应的多个反应室。 Klapperich 博士的实验室已利用 BioHelix 的等温扩增流程成功地在芯片中进行了核酸扩增。 Biohelix 的专有技术称为解旋酶依赖性扩增 (HDA)。它使用 DNA 解旋酶在 65:C 恒温的指数扩增过程中分离 DNA 链。与聚合酶链式反应 (PCR) 一样，HDA 检测使用竞争性内部对照（即掺入原始样品中的已知浓度的模板 DNA），可以通过与分析物相同的引物进行扩增，但单独检测；因此，我们能够在直接临床样本中检测扩增抑制剂。第一阶段的目标是： 1) 修改 BU 微流控装置设计，加入侧流条作为肉眼检测扩增产物的手段； 2) 证明使用目标 1 的设备执行集成测试的可行性，使用 1 mL 尿液中的 CT 和 104 的 NG 进行加标，为在包括侧流条的改进的 BU 芯片中执行的集成测定建立概念证明。我们第二阶段的具体目标是开发侧流读取器子系统，将其与 BU 仪器集成，构建预商业化的外观/工作原型，简化芯片设计和制造，以实现中试规模制造，并在 BioHelix / Quidel 实施一次性用品的中试规模生产。现阶段，Qiagen 是读者最有可能选择的制造商。在第二阶段结束时，我们应该为新检测系统的临床验证做好准备。