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.

这项小企业技术转移（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见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26500例住院治疗，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商和第二大禽肉出口国——必须尽量减少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对沙门氏菌微孔板检测的市场估计为2.4亿美元。该项目开发并实施了一种快速、简单和低成本的检测方法，可在15小时内进行沙门氏菌检测和定量，无需预富集步骤。它使加工商能够对加工环境中的沙门氏菌和生禽样品进行现场筛查，以更快地获得结果，从而更快地采取纠正行动，最大限度地减少沙门氏菌，并降低对消费者的下游风险。拟建项目将通过证明：对加工环境和生禽的纯培养物和样品的灵敏度（真阳性）和特异性（真阴性）≥90%，来证明多层、96孔微孔板检测和枚举沙门氏菌方法的可行性；15（±2）小时检测，无需预富集；稳定性：≥60天。该试验配方对7种沙门氏菌纯培养物和5种非靶菌的敏感性和特异性均≥95%，检测时间为15小时（±2）小时。本研究的主要技术挑战是在35种沙门氏菌生物和20种非靶标中实现所需的灵敏度、特异性、速度和稳定性，并使用各种靶标基质，以一种易于使用且具有成本效益的方法。控制非目标生物需要在微孔板的培养环境中评估选择性和差异性制剂，以及目标生物和非目标生物对营养物质的顺序和/或优先使用。将采用以下方法优化测试性能：(a)包容性测试；(b)排他性测试；(c)矩阵测试；(d)稳定性试验；(e)重复性测试。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。