SBIR Phase I: FRET-Aptamer Tests for Treated Wastewater

这个小企业创新研究（SBIR）第一阶段项目建议进一步发展其一步荧光共振能量转移(FRET)-DNA适体技术，用于快速（取样几分钟内得出分析结果），敏感，现场检测处理过的供水中的粪便细菌。操作技术公司（OpTech）之前成功开发了fret -适配体，该适配体结合了各种大肠杆菌菌株表面分子的共同核心抗原，并使用了fret -适配体的初步家族，使用商用现货（COTS）手持式和电池驱动的荧光传感器在水样中检测到每毫升10个细菌。这项技术的开发是为了满足联邦海滩法案的严格要求，但现在被提议作为一种更广泛的解决方案来验证处理过的废水的安全性。为了满足这一新要求，OpTech将为手持阅读器增加荧光活力测试，并与标准Colilert荧光测试结果进行比较。在第二阶段，OpTech将扩大其检测范围，包括其他肠道细菌、病毒和寄生虫。本研究的更广泛的影响是：1)提高了公众的饮用水安全；2)提高了污水处理设施的现场决策能力，以决定处理后的废水是否适合饮用、灌溉或排放回环境；3)在娱乐水域（海洋、湖泊甚至游泳池）中，更快地制定决策能力，以防止人类暴露于粪便污染。fret适体技术还应用于快速和便携式食品安全检测，国土安全和临床护理点诊断。

This Small Business Innovative Research (SBIR) Phase I project proposes to further develop its one-step fluorescence resonance energy transfer (FRET)-DNA aptamer technology for the rapid (assay results within minutes of sampling), sensitive, on-site detection of fecal bacteria in treated water supplies. Operational Technologies Corporation (OpTech) previously demonstrated successful development of FRET-aptamers that bind common core antigens of surface molecules from various strains of E. coli and used the preliminary family of FRET-aptamers to detect as few as 10 bacteria per milliliter in water samples with a commercial-off-the-shelf (COTS)handheld and battery-operated fluorescence sensor. The technology was in development to satisfy stringent requirements of the federal Beaches Act, but is now proposed as a broader solution for safety verification of treated wastewater. In order to meet this new requirement, OpTech will add a fluorescence viability test for the handheld reader and compare to standard Colilert fluorescence test results. In Phase II, OpTech will expand its repertoire of tests to include other enteric bacteria, viruses, and parasites.The broader impacts of this research are: 1) enhanced drinking water safety for the public 2)better on-site decision making ability for wastewater treatment facilities to decide if treated effluents are suitable for drinking, irrigation, or release back into the environment, 3) faster decision making ability at recreational waters (oceans, lakes, even swimming pools) with regard to potential closings to prevent human exposure to fecal contamination. FRET-aptamer technology also has applications in rapid and portable food safety testing, homeland security, and clinical point-of-care diagnostics.