Adaptation of enrichment-free Listeria diagnostic to food matrices

 描述(申请人提供)：食源性疾病每年摧毁数百万美国人的胃肠道微生物区系，通常导致生物失调，并增加其他疾病的风险，即肠易激综合征、肾功能衰竭、胰腺炎和糖尿病。此外，糖尿病患者患食源性疾病的风险更高，而且他们通常需要更长的时间才能康复。要了解食源性病原体对美国人口的影响有多大，可以考虑以下数字：每年有4800万人患病，2.8万人住院，至少3000人死亡。对医疗保健、商业和工业的影响也同样明显，导致每年的成本为140-160亿美元，包括直接医疗成本和因病损失的时间价值。在导致发病率和死亡率的食源性病原体中，细菌占大多数。关于这一点：细菌是导致疾病的前五大病原体中的四个，导致住院的前五个中的三个，以及前五个中的三个 会导致死亡。受污染的食品是这些疾病的原因，由于制造/包装厂、配送仓库或零售场所没有检测到，这些食品进入了公共供应。目前的检测是不够的，因为它们需要长时间的浓缩步骤，以降低假阴性和假阳性的可能性。这种浓缩导致向食品加工者提供可操作结果的巨大延误，他们只有两个选择之一：储存产品，直到检测结果出来，导致陈旧、质量较低的食品；或者在收到检测结果之前发货，增加消费公众的风险。Sample6正在开发世界上第一个无需浓缩的食源性病原体检测系统。检测/L是一种检测单个李斯特菌的快速筛选方法。在不到一个工作班次(&lt；7小时)的环境表面上的电池。我们专有的生物照明平台使这一快速转变成为可能。检测/L使食品制造商和包装商能够快速准确地检测和修复受污染的环境表面，并唯一保留有受污染风险的产品。然而，在生产、运输或供应链下游的批发和零售环境中，食品可能会受到内部污染。开发第二种检测模式至关重要，它可以像Detect/L对环境表面的检测那样快速和灵敏地检测食品。为此，拟议的第一阶段研究将采用Detect/L检测方法对成品进行检测，直接在五种特别容易受到李斯特菌污染的食品上检测李斯特菌：熟食火腿、烟熏三文鱼、墨西哥煎饼、菠菜和冰淇淋。首先，研究人员将确定食物对DETECT/L敏感度的抑制作用。接下来，将确定检测李斯特氏菌的金标法(美国农业部或美国食品药品监督管理局)的最低检测下限，然后将Detect/L与金标法进行面对面的比较。

 DESCRIPTION (provided by applicant): Foodborne illnesses devastate on the gastrointestinal microflora of millions of Americans each year, often resulting in dysbiosis and increase risk of other diseases, namely irritable bowel syndrome, kidney failure, pancreatitis and diabetes. Furthermore, diabetics are at increased risk of developing a foodborne illness, and they often take longer to recover. To realize the scale of the impact that foodborne pathogens have on the U.S. population, contemplate these figures: 48 million illnesses, 28,000 hospitalizations, and at least 3,000 deaths per year. The impact on healthcare, business and industry is no less pronounced, resulting in annual costs of $14-$16 billion, including direct medical costs and value of time lost to illness. Bacteria account for the majority of foodborne pathogens that cause morbidity and mortality. More to this point: bacteria are four of the top five pathogens that contribute to illness, three of the top five that cause hospitalizations, and three of the top five that cause mortality. Contaminated food is the cause of these illnesses, having entered the public supply due to a lack of detection at a manufacturing/packaging plant, distribution warehouse or retail location. Current tests are inadequate because they require a prolonged enrichment step to reduce the likelihood of false negatives and false positives. This enrichment causes tremendous delays in providing actionable results to the food processor, who are left with one of two choices: store the products until the test result comes back, resulting in older, lower-quality food; or shipping the food before receiving test results, putting the consuming public at increased risk. Sample6 is developing the world's first enrichment-free foodborne pathogen detection system. DETECT/L is a rapid screening assay that detects a single Listeria spp. cell on environmental surfaces in less than one work shift (<7h). Our proprietary Bioillumination Platform enables this expeditious turnaround. DETECT/L enables food manufacturers and packagers to quickly and accurately detect and remediate contaminated environmental surfaces, and solely retain product that is at risk of contamination. However, food can become contaminated internally during production, shipping, or in wholesale and retail environments downstream in the supply chain. It is of vital importance to develop a second testing modality that assays the foodstuff as rapidly and sensitively as DETECT/L does for environmental surfaces. To this end, the proposed Phase I study will adapt the DETECT/L assay for finished product testing, by directly detecting Listeria on five food products that are especially prone to Listeria contamination: deli ham, smoked salmon, queso fresco, spinach and ice cream. First, the researchers will determine the inhibitory effect that the food can have on DETECT/L's sensitivity. Next, the lower limit of detection (LLOD) of a gold-standard assay (USDA or FDA) in detecting Listeria will be determined, followed by a head-to-head comparison of DETECT/L with the gold-standard assays.