A fully integrated assay and platform for detecting Clostridium difficile.

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 peryear, 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. difficileto 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 thosechanges 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 canmechanically 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) ournovel 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.