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: 6783347
• 负责人: ROBERT James CARMAN
• 金额: $ 24.06万
• 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 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6783347
• 关键词: Clostridium difficile; Clostridium perfringens; 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）：顺磁颗粒（PMP）的使用及其在磁分析读取器（MAR）中的检测代表了一种新的“平台”技术，适用于侧流膜系统。在初步研究中，与使用免疫金或酶标记抗体的现有横向流动技术相比，新技术能够提供3到4个测井曲线的灵敏度。我们建议利用该技术开发一种新的艰难梭菌毒素a和B的快速横向流动系统。艰难梭菌是评估该技术的最佳候选者。这种厌氧病原体是工业国家院内腹泻和结肠炎的主要原因。尽管目前基于抗体的艰难梭菌检测比被认为是金标准的组织培养试验更快，但它们不能提供相同水平的灵敏度。在第一阶段，我们将优化开发艰难梭菌毒素PMP测试所需的条件。我们将优化PMP偶联工艺，用分离膜和捕集线工艺测试格式，评估稳定性，并开发粪便样品稀释剂。我们将开发一个原型装置，并开始研究以评估其性能特征。此外，我们将开发用于检测iota毒素的抗体，这是一种新发现的艰难梭菌毒素。二期将是一期的自然延伸，生产设备的技术将转移到GMP设施，我们将通过内部和现场研究评估GMP生产的设备。将收集和汇编数据，并准备510(k)文件。利用PMP横向流动技术平台，我们将在I期进行评估，我们还将开发一种诊断性艰难梭菌面板测试，这将有助于艰难梭菌疾病的流行病学和监测研究。此外，我们将开发抗生素相关性腹泻（AAD）的临床诊断面板测试，包括基于pmp的产气荚膜梭菌肠毒素测试，AAD的另一个原因。本项目开发的技术将广泛应用于开发新的高敏感粪便抗原检测。

项目成果

Molecular characterization and antimicrobial susceptibilities of extra-intestinal Clostridium difficile isolates.

肠外艰难梭菌分离株的分子特征和抗菌敏感性。

• DOI: 10.1016/j.anaerobe.2007.04.001
• 发表时间: 2007
• 期刊: Anaerobe
• 影响因子: 2.3
• 作者: Zheng,Limin;Citron,DianeM;Genheimer,ChristopherW;Sigmon,StephanieF;Carman,RobertJ;Lyerly,DavidM;Goldstein,EllieJC
• 通讯作者: Goldstein,EllieJC