Bacterial Pathogen Amplification & Real-Time Detection

描述(申请人提供)：众所周知，生物恐怖主义在美国是现实。然而，除了炭疽、鼠疫耶尔森氏菌和天花等难以安全生长和传播的A类制剂外，还有B类制剂可以用来感染我们的食物或水供应。这些生物包括细菌病原体、原生动物和病毒。除了这些天然的病原体外，还可以通过基因工程提高它们的毒力或抵抗标准的抗生素治疗。因此，迫切需要新的方法来快速检测食物和水传播的病原体，特别是如果它也可以用来鉴定这些微生物的药物敏感性的话。这个项目将开发这样的测试。 除了生物恐怖主义方面的担忧，食源性疾病还导致7600万人患病，其中包括每年在美国住院治疗的32.5万人和死亡的5000人。200多种已知疾病通过食物和饮料传播。传统的食源性病原体检测依赖于微生物学技术。AOAC推荐的这些标准培养方法需要5至7天才能完成。同时，产品被扣留等待发货，增加了存储成本，缩短了产品的保质期。就我们的供水而言，当发现问题时，公共卫生服务部门会发布警告，通常是在产品已经发布供公众使用之后。因此，迫切需要食品和水中病原体的快速检测方法来保护公众健康。 该计划的总体目标是开发一种集成的等温DNA扩增和检测系统，能够快速识别各种食物和水传播的病原体。所有NIAID生物防御B类食品和水传播的细菌病原体都将在该计划中检测到。提出了一种能够在封闭装置中等温放大和检测所有这些生物的单一集成系统。

DESCRIPTION (provided by applicant): As we all know, bio-terrorism in America is a reality. However in addition to the Category A agents like anthrax, Yersinia pestis and smallpox, which are difficult to safely grow and disseminate, exist the Category B agents that could be used to infect our food or water supply. These organisms include bacterial pathogens, protozoa, and viruses. In addition to these natural pathogenic organisms they could also be genetically engineered to increase their virulence or resist standard antibiotic treatments. Therefore new methods for rapid food and waterborne pathogen detection is greatly needed especially if it can also be used to identify drug sensitivity within these organisms. This program will develop such tests. In addition to bio-terrorism concerns, food-borne diseases cause 76-million illnesses, including 325,000 hospitalizations and 5,000 deaths in the US each year. More than 200 known diseases are transmitted through food and drink. Traditional detection of food-borne pathogens has relied on microbiologic techniques. These standard culture methods recommended by the AOAC require between 5 to 7 days to complete. Meanwhile, the product is held for shipment, increasing storage costs and reducing the product shelf life. In the case of our water supply, Public Health Service warnings are released when problems are identified, usually after the product is already released for public consumption. Consequently, rapid methods for food and water pathogen detection are greatly needed to protect the public health. The overall goal of this program is to develop an integrated isothermal DNA amplification and detection system capable of rapidly identifying a variety of food and waterborne pathogens. All of the NIAID Biodefense Category B food and waterborne bacterial pathogens will be detected in this program. A single integrated system capable of isothermal amplifying and detecting all of these organisms in a closed device is proposed.