Simultaneous Detection of Molecular and Serological Markers via Next-Generation Sequencing

Phase I SBIR proposals will be accepted. Fast-Track proposals will not be accepted. Phase I clinical trials will not be accepted. Number of anticipated awards: 1 Budget (total costs): Phase I: up to $243,500 for up to 6 months; Phase II of up to $1,000,000 and a Phase II duration of up to 2 years PROPOSALS THAT EXCEED THE BUDGET OR PROJECT DURATION LISTED ABOVE MAY NOT BE FUNDED. Background Simultaneous detection of serological markers including different antibody classes (IgM and IgG) and antigens in a multiplex fashion, as well as the characterization of molecular fingerprints of infectious agents is important for accurate diagnosis of several diseases. Proper identification of all necessary serological and molecular markers is of particular interest for outbreak investigations Page 130 and molecular surveillance. Thus, the development of platforms capable to simultaneously capture the required molecular and serological information is needed. Advanced characterization of a plethora of infectious agents relies on next generation sequencing (NGS) approaches, primarily using deep amplicon sequencing and Illumina sequencing technology. Cellular indexing of transcriptomes and epitopes by sequencing (CITE-Seq) allows next generation RNA sequencing as well as qualitative and quantitative analysis of proteins using capturing antibodies. CITE-Seq can be easily modified from single cell- to a bead-based approach for the specific detection of serological markers while simultaneously performing the conventional NGS protocols for genetic characterization. In combination, such methodologies could significantly improve the diagnosis for several diseases and syndromes including viral hepatitis. Project Goals Develop a multiplex NGS Illumina method for the simultaneous detection of viral hepatitis molecular and serological markers. Phase I Activities and Expected Deliverables 1. Create a standard operating procedure for antibody and antigen labeling. 2. Complete test runs on a MiSeq system to sequence viral hepatitis RNA and detect viral hepatitis serological markers. 3. Create a standard operating procedure for the complete molecular and serological laboratory detection of viral hepatitis. Impact Implementation of a NGS multiplex assay for the simultaneous detection of molecular and serological markers should significantly improve outbreak investigations, molecular surveillance and genetic relatedness studies for viral hepatitis and other infectious diseases. Commercialization Potential State laboratories are likely to benefit from implementing a multiplex approach for outbreak investigation and molecular surveillance of infectious agents able to capture both molecular and serological markers. Commercial labs will also likely benefit from implementing methodologies capable to characterize all serological and molecular markers for given infectious agents in a multiplex fashion.