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

这个小型企业创新研究(SBIR)第一阶段项目建议提高被广泛使用的检测和测量微量RNA(由细胞活跃地制造并包含在某些病毒中的遗传物质)方法的可靠性。目前大多数检测RNA的方法使用两种不同的酶；一种是将RNA转化为DNA，另一种是通过PCR扩增产生数十亿个拷贝。这种两种酶的方法存在严重的问题(例如，额外的操作、交叉污染、某些基因的不均匀扩增、不准确的扩增和低效率)。Lucigen公司已经发现了替代酶，并对其进行了表征，这些酶可以在单一反应中有效地执行RNA到DNA的转换和扩增。然而，这些方法需要得到更充分的发展和表征，以取代现有的方法。这些候选酶和我们收集的其他酶将被开发出来，并与第一阶段的可用酶进行比较。那些优于当前酶的酶将在第二阶段开发用于商业化。这项研究的更广泛影响包括大幅节省生物技术研究的成本，提高数据的准确性，以及提高诊断测试的可靠性。每年花在检测RNA靶标上的资金接近10亿美元，而且这个数字还在迅速增长。这种类型的研究严格依赖于可用的酶，并受到它们的不足的影响。这个SBIR项目将开发的改良酶有望极大地改善基础研究、诊断和药物开发，并将对人类健康、农业和微生物多样性研究产生重要影响。

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.