Adaptation of an enrichment-free Listeria diagnostic to food matrices

摘要 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 并增加其他疾病的风险，如肠易激综合征、肾功能衰竭、胰腺炎 还有糖尿病。此外，糖尿病患者患食源性疾病的风险增加，他们经常服用 恢复的时间更长。每年，美国每6个人中就有一个人因食源性病原体而患病。2014年，有 9起主要疫情导致19542例实验室确诊感染、4445例住院治疗和71例死亡 通常通过食物传播的病原体。李斯特菌是最严重的违法者之一，占96% 住院率和死亡率为12.9%。对医疗保健、商业和工业的影响也不亚于此 宣布，导致每年费用141亿至163亿美元，包括直接医疗费用和时间价值 因病失去了生命。此外，保持食品安全不受李斯特菌污染的成本估计在 24亿至26亿美元之间。被污染的食物是这些疾病的起因，已经进入公共供应。 由于制造/包装工厂、分销仓库或零售场所缺乏检测。当前 检测是不够的，因为它们需要长时间的浓缩步骤来降低错误的可能性 阴性和假阳性。这种浓缩导致在向 食品加工者，他们只有两个选择之一：储存产品，直到检测结果出来，结果 或在收到检测结果之前发运食品，使消费公众处于 风险增加。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。在更少的环境表面上的电池 超过一个工作班次(&lt；7小时)。Sample6‘S专有的生物照明平台使这种快速转变成为可能， 使食品制造商和包装商能够快速准确地检测和补救受污染的食品 环境表面，并只保留有污染风险的产品。然而，食物可以变成 在生产、运输或在下游的批发和零售环境中受到内部污染 供应链。开发另一种快速检测食品的检测方法至关重要 而且敏感，就像检测/L对环境表面所做的那样。为此，拟议的第二阶段研究将 继续第一阶段开始的工作，使DETECT/L检测方法适用于成品检测。首先， 研究人员将把李斯特菌的数量扩大到10个。下一步，将完成对 六种食物开始于第一阶段，另外还有30种食物。最后，Sample6将开发高通量方法，以便 食品制造商和包装商可以一次测试多个食品样本，以提高效率。

Abstract Foodborne illnesses devastate 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. Each year, 1 in 6 people in the U.S. are sickened by foodborne pathogens. In 2014, there were 19,542 laboratory confirmed infections, 4,445 hospitalizations and 71 deaths attributed to the 9 major pathogens transmitted commonly through food. Listeria is the one of the most serious offenders, with a 96% hospitalization rate and a 12.9% mortality rate. The impact on healthcare, business and industry is no less pronounced, resulting in annual costs of $14.1 to $16.3 billion, including direct medical costs and value of time lost to illness. Moreover, the cost of keeping food safe from Listeria contamination was estimated to range between $2.4-$2.6 billion. 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). Sample6’s proprietary Bioillumination Platform enables this expeditious turnaround, enabling 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 II study will continue the work begun in Phase I to adapt the DETECT/L assay for finished product testing. First, the researchers will expand the number of Listeria strains to 10. Next, assay development will be completed for the six foods begun in Phase I, plus 30 more foods. Finally, Sample6 will develop high-throughput methods so that food manufacturers and packagers can test multiple food samples at a time for efficiency.