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

该小企业创新研究（SBIR）一期项目提出改进实时PCR，一种基于dna的快速检测方法，用于检测沙门氏菌等食源性病原体。目前的方法不足以准确检测出巧克力、软奶酪和牛奶等对PCR有抑制作用的食品中的病原体。由于这些食物干扰了测定的灵敏度，因此给检测带来了挑战。这些食品需要更长时间、更耗费人力的检测，因为不准确的检测可能导致致命的假阴性。提出的改进将缩短和简化对这些食品的病原体检测。新的抗抑制突变酶和增强剂可以应用于PCR，并简化当前方案的其他阶段，如培养富集（病原体检测通常需要的潜伏期）和样品制备。这些改进可能还允许对更广泛的食品进行更快速、更准确的检测，而不仅仅是那些已知对PCR有抑制作用的食品。这项研究的更广泛影响包括更安全的食品供应，因为可以更快、更准确地检测食源性病原体。这将使食品生产商和制造商能够更快地对任何发现的污染作出反应，并促进立即召回受感染的食品。对于常规测试，这些创新将为测试实验室提供准确可靠的结果，同时节省他们的时间、劳动力和金钱。

This Small Business Innovation Research (SBIR) Phase I project proposes the improvement of real-time PCR, a DNA-based rapid-detection method, for detection of food-borne pathogens such as Salmonella. Current methods are inadequate to accurately detect pathogens in foods such as chocolate, soft-cheese, and milk, which are inhibitory to PCR. These foods present a challenge due to their interference with the sensitivity of the assay. Longer, more labor-intensive tests are required for these foods, as inaccurate detection can lead to potentially deadly false negatives. The proposed improvements will shorten and simplify pathogen detection for these foods. The novel inhibition-resistant mutant enzymes and enhancers can to applied to PCR, and simplify other stages of current protocols such as cultural enrichment (an incubation period usually required for pathogen detection) and sample preparation. These improvements may also allow for more rapid, accurate testing for a broad range of foods, not only those foods known to be inhibitory to PCR.The broader impacts of this research include a safer food supply, due to the faster, more accurate detection of food-borne pathogens. This will allow food producers and manufacturers to more quickly respond to any discovered contamination, and facilitate immediate recalls of infected food. For routine tests, these innovations will give testing labs accurate and reliable results, while saving them time, labor, and money.