SBIR Phase II: Inhibition-Resistant DNA Polymerases and Other Improvements for Detection of Food-borne Pathogens.

这个小型企业创新研究（SBIR）第二阶段项目提出了新的酶（DNA聚合酶）的开发和其他改进，用于通过DNA检测和扩增（PCR）快速检测食源性病原体。 PCR是一种非常快速和准确的病原体检测方法，通常在大约一天内给出结果，而不是通过培养方法生长和鉴定病原体所需的几天。 但有些食物，如巧克力、乳制品、肉类和香料，含有抑制PCR检测的成分。 目前在这些食品中进行快速病原体检测的策略包括长时间的培养富集步骤，然后稀释抑制剂和/或劳动密集型样品制备（DNA提取）以去除抑制剂。 抗抑制DNA聚合酶和食品特异性PCR增强剂代表了稀释或DNA提取的优雅，高科技替代品。 它们可以集成到现有的快速检测系统中，以促进对抑制性食品的快速准确检测。 这项研究的更广泛影响是减少了美国食源性疾病爆发的数量和严重程度，因为早期发现了食源性病原体。 更快、更准确地检测病原体将为食品制造商节省时间和金钱，并减少昂贵的产品召回需求。 这里开发的技术还可以扩展法医学的学科，在各种抑制剂的存在下恢复少量DNA是至关重要的，国防，快速检测用作武器的生物制剂可以挽救生命。

This Small Business Innovation Research (SBIR) Phase II project proposes the development of novel enzymes (DNA polymerases) and other improvements for rapid detection of food-borne pathogens by DNA detection and amplification (PCR). PCR is a very fast and accurate method of pathogen detection, typically giving results in about a day, instead of several days required to grow and identify pathogens by cultural methods. But some foods, such as chocolate, dairy products, meat, and spices, contain components that inhibit the PCR assay. Current strategies for rapid pathogen testing in these foods include long cultural enrichment steps followed by dilution of inhibitors and/or labor intensive sample preparation (DNA extraction) to remove inhibitors. Inhibition-resistant DNA polymerases and food-specific PCR enhancers represent elegant, high-tech alternatives to dilution or DNA extraction. They could be integrated into existing rapid-detection systems to facilitate rapid accurate testing in inhibitory foods. The broader impacts of this research are reducing the number and severity of outbreaks of food-borne illnesses in the United States due to early detection of food-borne pathogens. Faster, more accurate detection of pathogens will save time and money for food manufacturers, and reduce the need for costly product recalls. Technology developed here could also extend the disciplines of forensics, where recovery of small amounts of DNA in the presence of a variety of inhibitors is critical, and national defense, where rapid detection of biological agents used as weapons could save lives.