Method for Ultra-Sensitive Detection of Infectious Agents
Background The sensitive and accurate detection of spores and other infectious agents is essential in clinical environments and biodefense applications. Because of the excellent sensitivities obtained, polymerase chain reaction (PCR) methods are widely used for detection purposes. However, many difficulties are observed with present PCR methods, including poor specificity, high percentage of false positives due to poor specificity, and the requirement of multiple gene probes to confirm identification. Further, current PCR-based detection systems require complex cell lysis and nucleic acid extraction procedures before detection assays can be performed. Therefore, improved methods are needed to allow sensitive detection of infectious agents.
Invention Description We have linked the sensitivity of PCR with the selectivity of surface-based binding agents, such as antibodies or RNA aptamers, to generate an ultra-sensitive detection method applicable to a range of pathogens and cancer cells. No extraction steps are required, speeding assay time.
Antibodies or RNA aptamers adhere with great selectivity to cancer cells, virus capsids, or bacterial spores. Nucleic acid oligonucleotides attached to the binding agents are brought into proximity allowing sensitive real-time PCR amplification. Greatly improved sensitivity has been achieved through this amplification method over standard antibody-based assays, while maintaining virtually no background. Cancer cells can be detected in midst of 10,000-fold excess healthy cells. With some bacterial spores, as few as a single spore can be detected by this method.
Further, the use of antibody or other binding agents to select the pathogen cells allows multiple isolates and species to be detected, broadening the detection net and reducing the effects associated with genetic mutation. When adapting this approach to a new target assay, different antibodies can be linked to the same set of optimized oligonucleotides to speed product development cycle time. This methodology can be adapted for surface- or solution-based detection of bioterrorism threats and as a diagnostic tool for a range of disease-causing agents.
Reduction in false positives and false negatives Cell lysis or extraction not required Detection limits as low as one spore
Does not rely on genomic nucleic acids Can detect spores, viruses, or cancer cells via their surface antigens Can provide both static and dynamic readouts of the proximity of antigens on the cell surface
Market Potential/Applications This technology can be applied in several markets, including molecular diagnostics, biodefense applications, research and development, and environmental monitoring in hospital, biomanufacturing and other clean air settings.
IP Status One U.S. patent application filed
UT Researcher Andrew Ellington, Ph.D., Chemistry and Biochemistry, The University of Texas at Austin Supriya S. Pai, BA, Chemistry and Biochemistry, The University of Texas at Austin Matthew Levy, Ph.D., Chemistry and Biochemistry, The University of Texas at Austin
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