Rapid affordable aptamer-based diagnostic test for Hepatitis C virus Aptus Biosciences, LLC Bharat Gawande Summary Hepatitis C virus (HCV) is a global public health crisis, and an estimated 58 million people worldwide and 2 million people in the United States live with chronic HCV infection and are at risk of developing deadly liver cancer. U.S. Preventive Services Task Force recommends that all adults ages 18 to 79 be screened for HCV. Unfortunately, almost 75% of those do not know they are infected with HCV. Chronic HCV infection is entirely curable with a simple 12-week oral direct-acting anti-viral treatment; however, it needs a diagnosis. Although several anti-HCV antibody rapid HCV diagnostic tests are available, they can only report past infections, not current or chronic. Therefore, a second sensitive HCV RNA nucleic acid amplification test (NAT) is required to confirm active infection. However, NAT HCV infection tests are expensive, take time, are not available in many places such as rural or low-resource settings, and cannot be certainly performed at home. Therefore, to eliminate HCV from our population, which is the United States and the World Health Organization (WHO) mandate to at least 90% by 2030, we need a simple, easy-to-use, affordable, one-step rapid chronic HCV diagnostic test. The one-step rapid diagnostic test can be used at home for self-testing or at a clinic with low resources. In this SBIR Phase 1 proposal, Aptus Biosciences, LLC proposes to develop a rapid diagnostic test (RDT) for chronic HCV infection. The aptamer-based RDT developed at Aptus will detect HCV core antigen in the whole capillary blood, serum, or plasma samples. The HCV core antigen RDT will confirm the presence of active HCV infection and will probably be as good as the NAT HCV RNA test. Aptus Biosciences will use chemically modified aptamer selection technology to create highly sensitive and specific aptamers to detect HCV core antigen (Test line) and human IgG (Control line). We will deliver picomolar affinity aptamer sandwich pairs that bind to non-competing epitopes on targets to develop sandwich later flow assay (LFA). Our modified aptamer reagents are small (5- 10 kDa) that can reach the hidden epitopes on the target surface better than the large molecule, antibody-based reagents (150 kDa), ideally suited for sensitive and specific affordable rapid diagnostic tests. For our SBIR Phase 2 study, we will optimize aptamer sandwich pairs to improve sensitivity and assay performance, develop and validate prototype LFA with a small portable multi-use reader with Bluetooth and Wi-Fi connectivity to a Smartphone Mobile App (Android and iOS), and eventually submit a 510k application to the FDA for approval of rapid diagnostic HCV test for at home or point-of-care use. If successful, our rapid, one-step, affordable diagnostic test will save enormous healthcare spending on HCV liver disease management costs, improving the lives of millions of people and sig...