A Novel Diagnostic Test for Clostridium difficile-Associated Diarrhea (CDAD)

描述（由申请人提供）：本研究SBIR应用的目的是为全球重要但诊断不佳的医疗保健相关感染性疾病（即艰难梭菌相关腹泻或CDAD）的诊断检测开发范式转变。目前CDAD的实验室检测仅限于检测微生物和/或其毒素产物，无法区分感染患者和单纯定植患者，因此导致诊断不准确以及抗生素漏用/过度使用。为了克服这些限制，将使用Nanosphere的生物条形码检测技术开发一种超灵敏的检测方法，用于检测微生物和宿主对CDAD的反应目标。在第一阶段，将沿着粪便标本制备方案，开发针对毒素B（微生物靶标）和原肌球蛋白（宿主反应）的基于蛋白质和DNA的生物条形码检测。这种新方法的可行性将通过检测粪便样本中的CDAD来确定。在第二阶段，检测将进一步优化，以最大限度地提高灵敏度和特异性，仪器和软件将进行优化，以提供一个独立的“试剂盒”格式，（从样品接收到最终结果的完整测试在4小时内完成），可靠（灵敏度和特异性大于95%）、易于使用（使得经认证的实验室可以进行测定）和成本效益。 艰难梭菌相关性腹泻（CDAD）是一种全球重要但诊断不佳的卫生保健相关传染病。重要的是，这种疾病的病原体已成为一种重要的新兴病原体，在医院中发现了新的强毒株，社区获得性疾病的明显出现以及CDAD被认为是艾滋病毒感染者的关键病原体。一个更特异、更敏感的诊断平台将通过在非常早的时间点实现特异、正确的治疗来改善患者护理，从而降低严重疾病和死亡的潜在风险，并降低抗菌药物的过度使用。

DESCRIPTION (provided by applicant): The aim of this research SBIR application is to develop a paradigm shift in diagnostic testing for a globally important, yet poorly diagnosed, healthcare-associated infectious disease, namely Clostridium difficile-associated diarrhea, or CDAD. Current laboratory testing for CDAD is limited to detection of the organism and/or its toxin product(s) and does not differentiate infected from simply colonized patients, thus leading to inaccurate diagnosis as well as antibiotic miss/overuse. To overcome these limitations, an ultra-sensitive assay that detects both microbial and host response targets for CDAD will be developed using Nanosphere's bio-barcode assay technology. In phase I, protein and DNA-based Bio-barcode assays that target toxin B (microbial target) and tropomyosin (host-response) will be developed along with protocols for stool specimen preparation. The feasibility of this new approach will be established by testing for CDAD in stool samples. In phase II, the assays will be further optimized to maximize sensitivity and specificity, and instrumentation and software will be optimized to provide a stand-alone 'kit' format that is rapid (complete testing from specimen receipt to a final result in well under 4 hours), reliable (with sensitivity and specificity greater than 95%), easy to use (so that accredited laboratories can perform the assay), and cost efficient. Clostridium difficile-associated diarrhea, or CDAD, is a globally important, yet poorly diagnosed, healthcare-associated infectious disease. Importantly, the causative agent of this disease has become a crucial emerging pathogen with new virulent strains seen in the hospital, apparent emergence of community acquired disease and recognition of CDAD as a key pathogen for HIV-infected persons. A more specific and sensitive diagnostic platform will improve patient care by enabling specific, correct treatment at a very early time point, thus lowering the potential risk of severe disease and death as well as lower the overuse of antimicrobial agents.