Rapid, Single Multiplex Assay for Serum Pathogens

The goal of this project is to solicit the innovative development of diagnostic testing platforms that can simultaneously detect antigens of multiple pathogens and different antibody isotypes and/or immune factors against those pathogens in a single multiplex assay. The pathogens of interest include, but are not limited to, those with a high potential for causing global disease outbreaks that affect vulnerable populations (e.g., children, pregnant women, the elderly, etc.), and those targeted for global disease elimination or eradication. During implementation of the 6-month SBIR Phase 1 contract in 2019, Luna successfully constructed the sensing arrays of the MultiNanoSPRi multiplex assay, optimized the sensing chip surface activation system for human serum sample analysis, demonstrated precision tests and identified inter- and intra-assay coefficients of variabilities, identified limit of detections of ZIKV indicators RNA and IgG in serum, performed cross-reactivity test and multiplex detection among ZIKV indicators, demonstrated multiplex detection of 5 indicators: 3 for ZIKV (RNA, IgG and IgM) 1 for Ebola (IgG) and 1 for Malaria (antigen) spiked-in serum samples on a single chip, determined ZIKV antigen/antibody KD values and binding affinity, demonstrated detection of ZIKV indicators in clinical serum samples, and analyzed the manufacturing scalability and costs of MultiNanoSPRi. Luna submitted the SBIR Phase II proposal with four technical objectives aimed to further develop the MultiNanoSPRi assay. These objectives include detection of multiple pathogens, development of Luna SAS reagents and disposable gold-coated glass, development of quality control and algorithms for data output, and the validation of the system with frozen clinical samples. The identification and implementation of strategies for the validation of the MultiNanoSPRi platform for clinical diagnostic testing, as well as obtaining FDA regulatory approvals will be implemented in the Phase II contract. The envisioned product is MultiNanoSPRi assay kit with disposable chips ready to be used for commercially available SPRi instrument, such as OpenPlex (HORIBA Scientific). During the Phase II program, Luna will complete the MultiNanoSPRi development, demonstrate the use and stability of disposable gold-coated glass and Luna SAS reagents, and validate the assay platform and manufacturing of the MultiNanoSPRi kit for multiplex detection of ZIKV, CHIKV and DENV.