C. difficile Toxin Membrane Test with Magnetic Particles

使用磁性颗粒进行艰难梭菌毒素膜测试

• 批准号: 6689175
• 负责人: LIMIN ZHENG
• 金额: $ 22.38万
• 依托单位: TECHLAB, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6689175
• 关键词: Clostridium difficile; antibacterial antibody; artificial membranes; bacterial proteins; bacterial toxins; chickens; clinical research; diagnosis design /evaluation; diagnostic tests; diarrhea; enterotoxins; feces analysis; goats; human tissue; immunoconjugates; immunologic substance development /preparation; laboratory mouse; laboratory rabbit; magnetism; particle

DESCRIPTION (provided by applicant): The use of paramagnetic particles (PMP) and their detection in a magnetic assay reader (MAR) represent a new "platform" technology that is adaptable to lateral flow membrane systems. In preliminary studies, the new technology is capable of delivering 3 to 4 logs increased sensitivity over existing lateral flow technologies that use immunogold or enzyme-tagged antibodies. We propose to develop a new rapid lateral flow system for Clostridium difficile toxins A and B using this technology. C. difficile is an excellent candidate to evaluate the technology. This anaerobic pathogen is the leading cause of nosocomial diarrhea and colitis in industrialized countries. Although the current antibody-based tests for C. difficile are more rapid than the tissue culture assay, which is considered the gold standard, they do not offer the same level of sensitivity. In Phase I, we will optimize the conditions needed to develop a PMP test for the toxins of C. difficile. We will optimize the PMP conjugation process, test formatting with separation membrane and capture line processes, evaluate stability, and develop a fecal sample diluent. A prototype device will be developed and we will initiate studies to evaluate performance characteristics. In addition, we will develop antibodies for the detection of iota toxin, a newly recognized C. difficile toxin. Phase II will be a natural extension of Phase I. The technology for producing the device will be transferred to a GMP facility, and we will evaluate GMP produced devices through in-house and on-site studies. Data will be collected and compiled, and 510(k) documents will be prepared. Using the PMP lateral flow technology platform that we will evaluate in Phase I, we will also develop a diagnostic C. difficile panel test, which will be useful for epidemiological and surveillance studies of C. difficile disease. In addition, we will develop a clinical diagnostic panel test for antibiotic associated diarrhea (AAD), including a PMP-based test for C. perfringens enterotoxin, another cause of AAD. The technology developed in this project will be widely applicable for the development of new highly sensitive stool antigen tests.

描述（由申请人提供）：在磁性测定读取器（MAR）中使用顺磁性颗粒（PMP）及其检测代表了一种适用于侧流膜系统的新“平台”技术。 在初步研究中，新技术能够提供比现有的使用免疫金或酶标记抗体的侧向流技术增加3至4个对数的灵敏度。我们建议使用该技术开发一种新的用于艰难梭菌毒素A和B的快速侧向流动系统。C. difficile是评估该技术的绝佳候选者。这种厌氧病原体是工业化国家医院内腹泻和结肠炎的主要原因。虽然目前的C.艰难梭菌的检测比被认为是金标准的组织培养测定更快速，但它们不能提供相同水平的灵敏度。在第一阶段，我们将优化所需的条件，以开发用于C毒素的PMP测试。很难我们将优化PMP结合工艺，检测分离膜和捕获线工艺的格式化，评价稳定性，并开发粪便样本稀释剂。 我们将开发一个原型设备，并将启动研究，以评估性能特征。 此外，我们将开发用于检测iota毒素的抗体，iota毒素是一种新发现的C。艰难梭菌毒素 第二阶段将是第一阶段的自然延伸。生产器械的技术将转移到GMP设施，我们将通过内部和现场研究对GMP生产的器械进行评估。将收集和汇编数据，并编写510（k）文件。利用我们将在第一阶段评估的PMP横向流技术平台，我们还将开发诊断C。艰难梭菌平板试验，这将是有用的流行病学和监测研究的C。艰难病此外，我们将开发一种抗生素相关性腹泻（AAD）的临床诊断组合试验，包括基于PMP的C。产气荚膜杆菌肠毒素，AAD的另一个原因。本项目开发的技术将广泛应用于新型高灵敏度粪便抗原检测的开发。