SBIR-STTR Award

Immunofluorescent assays (IFA) are currently used primarily in support of diagnosis of patients with autoimmune disorders and infectious disease. Though it is the method of choice for many rheumatologists, Enzyme-Linked Immuno-sorbent Assays (ELISA) have become a more common immunologic procedure, in part due to cost considerations. Attempts to lower the cost of IFA by automating the readings are problematic due to poor photostability and high non-specific binding of the fluorescent dye. Recently, Hyperion, Inc. has introduced a system for automating preparations of IFA slides and has developed La Jolla BlueTM, a near-infrared-emitting, photostable dye with low nonspecific binding, that allows automation of all steps of the IFA. We have initiated studies with an automated digital instrument to acquire both fluorescence cellular images and quantitative readings of IFA. Experiments with the epifluorescence microscope have demonstrated nuclear fluorescence intensities with a coefficient of variation (CV) of <5 percent. The automated instrument will reduce labor costs for clinical IFA testing and enhance visual binding patterns with quantitative fluorescence intensity measurements. In Phase I, we will investigate feasibility in the key areas for development of the automated instrument. Phase II will proceed with instrument design and validation. PROPOSED COMMERCIAL APPLICATION: Testing with improved diagnostic sensitivity for autoimmune disorders, infectious diseases and various other clinical conditions. About 5,000 of the 25,000 laboratories in industrialized countries test for autoimmune diseases, purchasing pertinent reagent kits annually for $40 million in the US and $100 million worldwide. We expect to place 200, 400 and 600 reading instruments in the first three years, respectively, and to sell to the average user 200 reagent kits ($30,000) for autoimmune testing per year.

Diagnosis of auto-immune and connective tissue diseases such as Systemic Lupus Erythematosus, Sjorgren syndrome and scleroderma often makes use of an Immunofluorescent assay (IFA) for antinuclear antibodies (ANA). More detailed diagnostic support may also utilize an IFA test for double-stranded DNA (dsDNA). IFA tests detecting Anti-Neutrophil Cytoplasmic Autoantibodies (ANCA) are diagnostic for systemic necrotizing vasculitis and glomerulonephritis. All these IFA tests require significant laboratory preparation time from highly trained technicians using current, manual techniques. The VisiQuant system developed as a prototype in Phase I, combined with Hyperion's Hyprep automated assay preparation system will automate both preparation and reading of the IFA tests, achieving significant cost savings and potential error reduction. During the Phase II project period, an updated VisiQuant Microscope will be introduced along with improved ANA assay and preparation procedure. The assay will be improved by applying results of a study of assay parameters and conditions. The Microscope will be upgraded by incorporating engineering enhancements to the optics and electronics, including features to optimize the joint operation of the Microscope with the HyPrep, and software improvements to the fluorescence intensity algorithm. New test kits for dsDNA based on Crithidia luciliae and for ANCA will be developed, tested and brought to market. Development work in automated image classification will lead to a complementary product that finds the best match to each sample image from a standard ANA image pattern library. Further development of the image classification software will be done based on input from the consultants. This will lead to an advanced Image Classification system to be brought to market at the end of the Phase II period. Image classification to support ANCA test will be started but is not expected to be ready for market until after completion of this project. These developments will result in Hyperion obtaining more than 18% share of the global market for autoimmune IFA based IVD tests by 2010. The development team includes personnel with many years of experience in dye chemistry, immunology in vitro device development, instrument development and image processing.

Thesaurus Terms:
biomedical automation, biomedical equipment development, fluorescence microscopy, imaging /visualization /scanning, immunofluorescence technique DNA, antinuclear autoantibody bioimaging /biomedical imaging, clinical research, fluorescent dye /probe, human tissue