The objective of this project is to transition Argonne-developed intellectual property into the commercial marketplace, and develop a protein array for clinical surveillance and diagnosis of common respiratory infections, including influenza A, influenza B, parainfluenza (types 1-3), human respiratory syncitial virus (RSV), adenovirus (types A-f), SARS coronavirus, group A Streptococcus, group B Streptococcus, Mycobacterium tuberculosis, Mycoplasma pneumonia, Haemophilus influenza, and Staphylococcus pneumoniae (12 pathogens). An array-based approach to respiratory disease diagnosis is recommended because it is difficult to differentiate between viral and bacterial infections based on clinical signs or radiology of the presenting patient, and because single-antibody tests are notoriously prone to false positives. A competitive immunoassay is proposed to militate against detector antibody cross-reactivity in a sandwich immunoassay array format; the sandwich immunoassay is reserved as a contingency to achieve the Phase 1 milestones. The decision logic and results from the protein array will enable public health officials to rapidly identify infected persons and intelligently administer costly and/or limited quantities of antibiotics or anti-viral therapies either in the clinic or in the field within 20 min of sample acquisition. We will meet these objectives by taking advantage of Akonni's exclusively-licensed intellectual property portfolio acquired from Argonne National Laboratory; proprietary antibodies, sample preparation reagents and know-how from Quidel Corporation; protein array expertise at Argonne National Laboratory; and clinical practice at Little Company of Mary Hospital.
Specific Aims for the Phase 1 project are to: develop and implement a sample preparation and multiplexed immunoassay procedure within Akonni's fluidic development model sub-circuits; determine cross- reactivity of target antigens with Quidel antibodies in the competitive protein array; determine limits of detection and cross-reactivity for 12 individual pathogens in phosphate buffered saline and amended nasal swabs, lavage or sputum; and define process flow and instrument requirements for integrated fluidic cartridge development, production and testing in Phase 2. Successful completion of the project Milestones will provide sufficient justification to proceed with a Phase 2 proposal, where the Specific Aims will be to: integrate the sample preparation and protein array functions into a self-contained, integrated, injection-molded test cartridge; test and evaluate reagent stability and longevity within the test cartridge; encode the array decision logic into user-friendly software; test and evaluate the integrated prototype on neat and amended nasal swabs, throat swabs, nasal lavage and/or sputum clinical samples in accordance with 510K submission guidelines; and begin cartridge production scale-up in collaboration with Quidel. ? ? The Diagnostic Respiratory Protein Array test cartridge will represent a low-cost, simple diagnostic tool for identifying individuals infected with respiratory pathogens. The underlying platform developed herein will likewise find broad application in many areas of disease surveillance, with an emphasis on point-of-use applications. ? ? ? ? ?
