STTR Phase I: Feasibility of a two-phase test tube assay for rapid detection and enumeration of Vibrio vulnificus and Vibrio parahaemolyticus in oysters

STTR 第一阶段：用于快速检测和计数牡蛎中创伤弧菌和副溶血弧菌的两相试管测定的可行性

• 批准号: 1819545
• 负责人: Chris Ramezanpour
• 金额: $ 22.5万
• 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: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1819545&HistoricalAwards=false
• 关键词: STTR; Phase; Feasibility; two; phase

The broader impact/commercial potential of this Small Business Technology Transfer (STTR) project will be the development of a detection assay to reduce the incidence of Vibrio-related infections. Infections from Vibrio parahaemolyticus and Vibrio vulnificus are the leading causes of seafood-borne infections and fatalities, respectively, and stem primarily from raw oysters. Development and implementation of rapid, accurate, inexpensive and easy-to-use testing methods would expand Vibrio data collection and enhance preventive measures by state shellfish labs (monitor harvest areas), and oyster processors and post-harvest processors (quality control at facilities). The assays will be available to users in simple, ready-to-prepare kits. These kits will make it easy for non-specialized personnel to test oyster lots still under refrigeration without disrupting logistics, and help U.S. oyster processors and post-harvest processors improve risk management tools and access export markets with restrictions on Vibrio levels. The tests also will improve environmental risk evaluations for harvest areas by state shellfish labs, FDA, National Oceanic and Atmospheric Administration, and other members of the National Shellfish Sanitation Program. The tests will be marketed in the U.S., and other large oyster producing nations.This STTR Phase I project proposes to develop two-phase, thermostable test tube media formulations for the detection and enumeration of pathogenic V. parahaemolyticus, total V. parahaemolyticus, and V. vulnificus in oysters. The goal is to demonstrate a) 95% sensitivity and specificity of each test with pure cultures and oyster samples; b) 24-hr detection time; c) ease-of-use of the ready-to-prepare kits, based on time, steps, equipment, materials and feedback from industry; and d) low cost. The main technical hurdles are: 1) Increasing formulation volumes for test tube assays since linear increases of agar layer volumes in test tube media formulations could impact Vibrio organism motility, and consequently detection speed and accuracy; and 2) replacing a non-heat stable antibiotic component, Polymixin B of the V. vulnificus test, with a heat stable alternative, like ampicillin or lincomycin, to improve ease-of-use.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) 项目更广泛的影响/商业潜力将是开发一种检测方法，以减少弧菌相关感染的发生率。副溶血弧菌和创伤弧菌感染分别是海鲜传播感染和死亡的主要原因，并且主要源于生牡蛎。开发和实施快速、准确、廉价且易于使用的检测方法将扩大弧菌数据收集，并加强国家贝类实验室（监测收获区域）以及牡蛎加工商和收获后加工商（设施的质量控制）的预防措施。这些检测将以简单、可随时准备的试剂盒形式提供给用户。这些套件将使非专业人员能够轻松测试仍在冷藏状态下的牡蛎批次，而不会中断物流，并帮助美国牡蛎加工商和收获后加工商改进风险管理工具并进入限制弧菌水平的出口市场。这些测试还将改善国家贝类实验室、FDA、国家海洋和大气管理局以及国家贝类卫生计划其他成员对收获区的环境风险评估。这些测试将在美国和其他大型牡蛎生产国销售。该 STTR 第一阶段项目建议开发两相、耐热试管培养基配方，用于检测和计数致病性副溶血弧菌、总副溶血弧菌和牡蛎中的创伤弧菌。目标是证明 a) 使用纯培养物和牡蛎样本进行的每次测试具有 95% 的灵敏度和特异性； b) 24小时检测时间； c) 根据时间、步骤、设备、材料和行业反馈，即用型套件的易用性； d) 成本低。主要技术障碍是： 1) 增加试管测定的配方体积，因为试管培养基配方中琼脂层体积的线性增加可能会影响弧菌微生物的运动性，从而影响检测速度和准确性； 2) 用热稳定替代品（如氨苄西林或林可霉素）取代非热稳定抗生素成分（创伤弧菌测试中的多粘菌素 B），以提高易用性。该奖项反映了 NSF 的法定使命，并被视为值得通过使用基金会的智力优点和更广泛的影响审查标准进行评估来支持。