Rapid, Handheld, Point-of-Care Stool Sample-prep Device for the Extraction and Detection of Nucleic Acid from Diarrheal Pathogens

Abstract Diarrheal disease is a major health issue worldwide resulting in nearly 500,000 deaths annually in children under 5 years of age. Those who survive have higher risk of stunted growth and cognitive development. Identification of the gastrointestinal (GI) pathogens that cause diarrheal disease helps guide effective treatment. Currently, in the developed world, this is done using multiplexed PCR laboratory tests which are expensive, slow and require a sophisticated high complexity laboratory. Unfortunately, such laboratory tests are not available in remote and resource limited regions in the developing world where mortality due to diarrhea is the highest. Thus, we propose to develop an integrated, low-cost, point-of-care (POC) test to identify the GI pathogens that cause diarrheal diseases. This test will be useful in resource-limited settings of the developing world as well as in primary care and urgent care settings of the developed world where it will be useful to detect the GI pathogens in real time, in the clinician's office so the patient can be treated with the right drugs, right away, without loss of patients to follow up. The major problem with detecting gastrointestinal (GI) pathogens is the complexity of stool samples, often filled with particulate and molecules such as bile salts. Such contaminants can inhibit signal amplification and detection if not properly removed from the sample. In previous SBIR projects, GoDx has developed sample preparation chemistry, DNA amplification and detection assays on lateral flow paper strips for a suite of GI pathogens. Our next step is to develop a pathogen detection device that fully integrates sample processing and signal amplification, which can be used rapidly at the point of care (POC). To achieve this, we propose to develop a stool sample prep device that can robustly provide viable DNA for amplification and detection all while requiring minimal user action to do so. The technology proposed here will be able to extract and enrich DNA from a swab of stool in a matter of minutes without the need for centrifuges or pumps. In Phase 2 of this project, we will fully integrate the GoDx’s amplification and detection assays with the sample prep technology developed here, providing a seamless, minimal step POC device for the detection of GI pathogens that cause diarrheal diseases around the world.