SBIR Phase I: Thermostable Reverse Transcriptases for Single Enzyme RT-PCR

This Small Business Innovation Research (SBIR) Phase I project proposes to improve the reliability of widely used methods for detecting and measuring trace amounts of RNA (genetic material actively made by cells and contained within certain viruses). Most current methods for detecting RNA use two different enzymes; one to convert RNA to DNA and a second to generate billions of copies by PCR amplification. This two enzyme approach has serious problems (e.g., additional manipulations, cross contamination, uneven amplification of certain genes, inaccurate amplification, and low efficiency). Alternative enzymes have been discovered and characterized at Lucigen that efficiently perform both conversion of RNA to DNA and amplification in a single reaction. However, these need to be more fully developed and characterized to replace existing methods. These candidates and others in our collection will be developed and compared to available enzymes in Phase I. Those that are superior to current enzymes will be developed for commercialization in Phase II. The broader impacts of this research include substantial cost savings in biotechnology research, increased accuracy of data, and improved reliability of diagnostic tests. Nearly $1 billion is spent annually on detecting RNA targets, and this amount is growing rapidly. This type of research is strictly dependent on the available enzymes and is compromised by their deficiencies. Improved enzymes to be developed in this SBIR project promise to greatly improve basic research, diagnostics, and drug development, and will have an important impact on human health, agriculture, and research in microbial diversity.