A fully integrated assay and platform for detecting Clostridium difficile.

描述(由申请人提供)：这一拟议项目的主要目标是开发一种从人的粪便中检测艰难梭状芽胞杆菌的即时诊断分析和平台。另外两个目标是检测艰难梭菌B1/NAP/027的超强毒力菌株，以及检测医疗器械拭子中的艰难梭菌孢子。艰难梭菌相关性疾病(CDAD)现在是美国最常见的医疗获得性感染，估计每年有300万CDAD病例，每年造成10-32亿美元的损失。在住院时间长达两周的患者中，艰难梭菌的获得率估计为13%，在住院时间超过四周的患者中，获得率为50%。将艰难梭菌的检测纳入护理地点设置，将有助于患者更及时地接受治疗，并降低疾病传播的风险。监测艰难梭菌表面污染的能力是该项目的独特贡献。全国和全球的医院、疗养院和康复机构将欢迎能够监测艰难梭菌及其孢子对环境污染的负担，采取相应的感染控制程序，并核实这些变化已达到预期结果。克莱蒙特生物解决方案公司(CBio)正在开发一种集成的一次性分析设备和平台，该设备和平台基于我们用于样品制备的专有一次性墨盒PureLyse(R)的组合，该墨盒可以在不到5分钟的时间内机械裂解细胞并提取DNA，并带有快速等温核酸放大系统，两者都嵌入了一次性设备中。该平台提供对扩增产物的检测，并基于CBio的便携式荧光仪。PureLyse(R)室将安装在我们新型的OmniValve中，以创建一种有助于快速高效制造的分析设备。 公共卫生相关性：艰难梭菌最近已成为美国最常见的医院获得性感染。我们建议开发一种用于从人的粪便和医疗设备的拭子中检测艰难梭菌的点式分析和便携式平台，其基础是：(1)我们的新型PureLyse(R)快速和一次性样品制备系统，该系统可以在不到5分钟的时间内裂解细胞并提取DNA；(2)DNA的快速等温放大；以及(3)用我们的FluoriSense(R)便携式荧光仪进行荧光检测。

DESCRIPTION (provided by applicant): The primary goal of this proposed project is to develop a point-of-care diagnostic assay and platform for the detection of Clostridium difficile from human stool. Two additional goals are to detect the hypervirulent strain of C. difficile B1/NAP/027 and to detect C. difficile spores from swabs of medical devices. C. difficile associated disease (CDAD) is now the most common healthcare acquired infection in the United States with an estimated three million CDAD cases per year, costing $1- 3.2 billion per year. The frequency of C. difficile acquisition is estimated to be 13% in patients with hospital stays of up to two weeks, and 50% in those with hospital stays longer than four weeks5. Bringing detection of C. difficile to the point-of-care setting will facilitate more timely treatent for patient, as well as reduce the risk of spread of the disease. The capability to monitor surfaces for C. difficile contamination is a unique contribution of this project. Hospitals, nursin homes, and rehabilitation facilities, both nationally and globally, will welcome the ability to monitor the burden of environmental contamination by C. difficile and its spores, respond with infection control procedures accordingly, and verify that those changes have achieved the desired result. Claremont BioSolutions (CBio) is developing an integrated disposable assay device and platform that is based on the combination of our proprietary disposable cartridge for sample preparation, PureLyse(R), that can mechanically lyse cells and extract DNA in less than five minutes, with a rapid isothermal nucleic acid amplification system, both embedded into a disposable device. The platform provides detection of the amplification product and is based on CBio's portable FluoriSense" fluorometer. The PureLyse(R) chamber will be mounted within our novel OmniValve" to create an assay device that lends itself to rapid and efficient manufacturing. PUBLIC HEALTH RELEVANCE: Clostridium difficile has recently become the most common hospital-acquired infection in the United States. We propose to develop a point-of-care assay and portable platform for detecting Clostridium difficile from human stool and from swabs of medical devices based on the combination of (1) our novel PureLyse(R) rapid and disposable sample preparation system that can lyse cells and extract DNA in less than five minutes, (2) rapid isothermal amplification of DNA, and (3) fluorescence detection with our FluoriSense(R) portable fluorometer.