Food safe pathogen surrogates based on encapsulated short DNA sequences for rapid verification of sanitation in food processing

食品安全病原菌代用品用于食品加工卫生的快速验证 第二阶段 项目摘要 食品安全是一个影响全球的重大问题。疾控中心估计，仅在美国， 每年有3000人死于食源性疾病。据估计，总共有76 2013年发生了100万例与食品有关的疾病，美国在治疗上花费了400亿美元 而不是预防(《食品质量和安全杂志》，2015)。即使是食源性的 疾病通常归因于肉类、乳制品和海鲜，这些疾病是由于受污染的农产品而爆发的。 一直呈上升趋势(Harris等人，2003年；Lynch等人，2009年；Callejón等人，2015年)。 新鲜水果和蔬菜大多是在户外种植的，因此很容易受到 潜在的环境污染。洗涤是加工者使用的主要方法 以确保他们供应的农产品的安全。然而，洗涤并不总是有效的 去除微生物污染物(去污)。如果洗涤系统管理不当， 交叉污染可以将一个小的污染事件转变为暴发事件。 SafeTraces正在提议开发SafeTracers™，一种可靠的、可重复的、 对成品进行持续、快速的现场测试，以确保适当的清洗 评估新鲜农产品的去污和交叉污染风险。安全跟踪程序™是 可食用(通常被认为是安全的)微粒，标记有无生命、无活性的DNA， 在洗涤条件下表现得像病原体。它们将启用验证、验证和 持续监测洗涤系统中的去污和交叉污染。 SafeTracers™可以很容易地在任何洗涤系统上进行改装，并且坚固耐用，操作简单 做手术吧。 在第二阶段，我们将比较安全示踪剂™与主要病原体的 非生物材料(玻璃和不锈钢)及其他材料的附着、分离、转移和失活 制作(菠菜)表面。我们将开发一个简单的工作流程来采样成品， 测量SafeTracers的™水平，并评估洗涤过程的有效性，所有这些都在 30分钟。此外，我们将开发一种基于SafeTracers™的方法来评估E. 在一次性洗涤系统和水槽洗涤系统中，叶类农产品中的大肠杆菌交叉污染 使用氯化水洗。最后，我们将完成对两者的验证研究 SafeTracers基于™的方法，旨在评估未充分净化和 在经过验证的条件下运行的洗涤系统和加工过程中的交叉污染 绿党。

Food Safe Pathogen Surrogates for Rapid Verification of Sanitation in Food Processing Phase II Project Summary Food safety is a major issue with global impact. The CDC estimates that in the U.S. alone, 3,000 deaths are caused each year due to foodborne illnesses. It is estimated that a total of 76 million food-related illness cases occurred in 2013, with the U.S. spending $40 billion on treatment rather than prevention (Food Quality and Safety Magazine, 2015). Even though foodborne illnesses are often attributed to meat, dairy, and seafood, outbreaks due to contaminated produce have been on the rise (Harris et al., 2003; Lynch et al., 2009; Callejòn et al., 2015). Fresh fruits and vegetables are mostly grown outdoors, leaving them susceptible to potential environmental contamination. Washing is the primary method employed by processors to ensure the safety of the produce they supply. However, washing is not always effective in removing microbial contaminants (decontamination). If the wash system is improperly managed, cross contamination can covert a small contamination event into an outbreak event. SafeTraces is proposing the development of SafeTracers™, a reliable, repeatable, continuous, fast, on-site test performed on finished product, to ensure appropriate washing by assessing decontamination and risk of cross-contamination of fresh produce. SafeTracers™ are edible (Generally Recognized As Safe) microparticles, tagged with non-living, non-viable DNA, that behave like pathogens under wash conditions. They will enable validation, verification, and continuous monitoring of decontamination and cross-contamination in wash systems. SafeTracers™ will be easily retrofitted on any wash system, and will be robust and simple to operate. During Phase II, we will compare SafeTracers™ to prominent pathogens for their attachment, detachment, transfer, and inactivation on abiotic (glass and stainless steel) and on produce (spinach) surfaces. We will develop a simple workflow to sample finished product, measure SafeTracers™ levels, and assess the effectiveness of the wash process, all in less than 30 minutes. Furthermore, we will develop a SafeTracers™-based method to assess the risk of E. coli cross contamination for leafy produce commodities in both one-pass and flume wash systems using chlorinated water wash. Finally, we will complete validation studies for the two SafeTracers™-based methods aimed at assessing the risks for insufficient decontamination and cross contamination in wash systems operating under validated conditions and processing leafy greens.