STTR Phase I: Feasibility of multi-layer microplate test for rapid detection and enumeration of Salmonella spp. in raw poultry and processing environment samples

这个小企业技术转让(STTR)第一阶段项目的更广泛影响将是减少沙门氏菌的感染和暴发，沙门氏菌是美国报告的最常见的食源性疾病的细菌原因之一。沙门氏菌每年导致超过135万人患病，26500人住院，其中20%的感染可归因于家禽。鉴于沙门氏菌疾病的数量在过去十年中增加了约30%，美国家禽业-世界上最大的禽肉生产国和第二大禽肉出口国-必须将沙门氏菌感染的风险降至最低。美国家禽加工商和相关卫生服务提供商的沙门氏菌微孔板检测市场估计为240美元。该项目开发和实施了一种快速、简单和低成本的沙门氏菌检测和量化测试，无需预浓缩步骤，只需15小时。它使加工人员能够在加工环境和生禽肉样本上进行现场沙门氏菌筛查，以更快地获得结果，从而能够更快地采取纠正行动，将沙门氏菌降至最低，并降低消费者面临的下游风险。拟议的项目将展示用于沙门氏菌检测和计数的多层、96孔微孔板测试方法的可行性，展示：≥对纯培养物和加工环境和生禽肉样本的灵敏度(真阳性)和特异性(真阴性)为90%；检测在15(±2)小时内完成，无需预浓缩；以及稳定性：≥60天。该试验基础配方对7株沙门氏菌纯培养物和5株非靶标微生物具有95%的灵敏度和特异度，检测时间为15小时(±2)小时。这项研究的主要技术挑战是以一种易于使用和具有成本效益的方法，在35种沙门氏菌生物和20种非靶标生物中，以一种易于使用和具有成本效益的方法，实现所需的灵敏度、特异度、速度和稳定性。对非目标生物的控制需要评估选择性和差别剂，以及目标生物和非目标生物在微孔板培养环境中对营养物质的顺序和/或优先使用。以下方法将用于优化测试性能：(A)包容性测试；(B)排他性测试；(C)矩阵测试；(D)稳定性测试；(E)重复性测试。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact of this Small Business Technology Transfer (STTR) Phase I project will be reduced incidence of infections and outbreaks from Salmonella, one of the most frequently reported bacterial causes of food-borne illness in the US. Each year Salmonella causes over 1.35 M illnesses and 26,500 hospitalizations, with 20 percent of infections attributable to poultry. Given that the number of Salmonella illnesses has increased about 30 percent over the past decade, the US poultry industry - the world's largest producer and the second largest exporter of poultry meat - must minimize the risk of Salmonella infection. The market for the Salmonella microplate test with US poultry processors and the associated sanitation service providers is estimated at $240 M.This project develops and implements a rapid, simple and low-cost test for Salmonella detection and quantification in 15 hours with no pre-enrichment step. It enables processors to run on-site screening for Salmonella in the processing environment and on raw poultry samples to get faster results, enabling faster corrective action to minimize Salmonella and reducing downstream risks to consumers. The proposed project will show the feasibility of a multi-layer, 96-well microplate test method for Salmonella detection and enumeration by demonstrating: ≥ 90% sensitivity (true positive) and specificity (true negative) with pure cultures and samples of the processing environment and raw poultry; detection in 15 (± 2) hours, with no pre-enrichment; and stability: ≥ 60 days. The test base formulation has shown ≥ 95% sensitivity and specificity with 7 pure cultures of Salmonella and 5 non-target organisms, and detection in 15 hours (± 2) hours. The main technical challenges of this research are achieving desired sensitivity, specificity, speed and stability across 35 Salmonella organisms and 20 non-targets, with a variety of target matrices, in a method easy to use and cost-effective. Controlling for non-target organisms requires evaluation of selective and differential agents as well as sequential and/or preferred use of nutrients by target and non-target organisms in the culturing environment of the microplate. The following methods will be used to optimize test performance: (a) Inclusivity testing; (b) Exclusivity testing; (c) Matrix testing; (d) Stability testing; and (e) Reproducibility testing.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.