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

STTR 第一阶段：用于快速检测和计数沙门氏菌的多层微孔板测试的可行性。

• 批准号: 2135699
• 负责人: Chris Ramezanpour
• 金额: $ 25.6万
• 依托单位: Secure Food Solutions
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2135699&HistoricalAwards=false
• 关键词: STTR; Phase; Feasibility; multi; layer

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.

该小企业技术转让 (STTR) 第一阶段项目的更广泛影响将减少沙门氏菌感染和爆发的发生率，沙门氏菌是美国最常报告的食源性疾病细菌原因之一。沙门氏菌每年导致超过 135 万例疾病和 26,500 例住院治疗，其中 20% 的感染归因于家禽。鉴于沙门氏菌疾病的数量在过去十年中增加了约 30%，作为全球最大的禽肉生产国和第二大禽肉出口国的美国家禽业必须最大限度地降低沙门氏菌感染的风险。美国家禽加工商和相关卫生服务提供商进行沙门氏菌微孔板测试的市场估计为 2.4 亿美元。该项目开发并实施了一种快速、简单且低成本的沙门氏菌检测和定量测试，可在 15 小时内进行，无需预富集步骤。它使加工商能够在加工环境和生家禽样品上进行沙门氏菌现场筛查，以更快地获得结果，从而更快地采取纠正措施，最大限度地减少沙门氏菌并降低消费者面临的下游风险。拟议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，具体方法是：纯培养物以及加工环境和生家禽样品的灵敏度（真阳性）≥ 90% 和特异性（真阴性）； 15（±2）小时内检测，无需预富集；稳定性：≥60天。测试基础配方对沙门氏菌和 5 种非目标生物的 7 种纯培养物显示出 ≥ 95% 的灵敏度和特异性，并在 15 小时 (± 2) 小时内进行检测。这项研究的主要技术挑战是通过易于使用且经济高效的方法，利用各种目标基质，在 35 种沙门氏菌生物体和 20 种非目标物中实现所需的灵敏度、特异性、速度和稳定性。控制非目标生物体需要评估选择性和差异性试剂，以及目标生物体和非目标生物体在微孔板培养环境中顺序和/或优选使用营养物。将使用以下方法来优化测试性能： (a) 包容性测试； (b) 排他性测试； (c) 矩阵测试； (d) 稳定性测试； (e) 再现性测试。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。