Bacterial Pathogen Amplification & Real-Time Detection

细菌病原体扩增

• 批准号: 6640955
• 负责人: JACK R. U'REN
• 金额: $ 9.99万
• 依托单位: SAIGENE CORPORATION
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2003-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6640955
• 关键词: Campylobacter; Escherichia coli; Listeria; Salmonella; Salmonella food poisoning; Shigella; Vibrio cholerae; Yersinia pestis; bacterial food poisoning; biohazard detection; bioterrorism /chemical warfare; food contamination; food processing /preparation; nucleic acid amplification techniques; nucleic acid probes; nucleic acid quantitation /detection; water sampling /testing; water supply

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 to resist standard antibiotic treatments. Therefore new methods for rapid sensitive food and waterborne pathogen detection are greatly needed, especially if they can also be used to identify drug sensitivity within these organisms. Bio-terrorism using a food pathogen is not just a hypothetical threat to America. A religious cult in Dalles Oregon sickened at least 751 people by contaminating food in grocery stores and restaurants in the fall of 1984. The group simply grew cultures of the food pathogen Salmonella typhimurium that they obtained from their local scientific supply house and sprinkled the cultures on produce in the grocery stores and the restaurant salad bars. If the group had used a more deadly pathogen like Salmonella typhi that causes typhoid fever many people would certainly have died. The overall goal of this program is to develop an integrated isothermal DNA amplification and a probe array detection slide capable of rapidly identifying a variety of food and waterborne pathogens. All of the NIAID Biodefense Category B food and waterborne bacterial pathogens E. coli, Vibrio cholera, Shigella dysentery, Salmonella species, Listeria monocytogenes, Camphylobacter jejuni, and Yersinia enterocolitica will be detected in this program. A single integrated slide capable of isothermal amplifying and detecting all of these organisms in real-time in a closed sealed device is proposed. The program can also distinguish live organisms from dead organisms killed by the food or water sanitation process. Also, the test can be used to identify the antibiotic sensitivity of the pathogen to identify genetically altered organisms.

描述（由申请人提供）：众所周知，美国的生物恐怖主义是现实。然而，除了A类A类药物之外，诸如炭疽病，耶尔森氏菌和天花很难安全地生长和传播，还存在可用于感染我们的食物或供水的B类药物。这些生物包括细菌病原体，原生动物和病毒。除了这些天然的致病生物外，它们也可以在基因上进行基因设计，以增加其毒力或抵抗标准的抗生素治疗。因此，非常需要新的快速敏感食品和水传播病原体检测的方法，尤其是如果它们也可以用来鉴定这些生物体内的药物敏感性。使用食品病原体的生物恐怖不仅是对美国的假设威胁。 1984年秋天，俄勒冈州达勒斯（Dalles）的一个宗教邪教在杂货店和餐馆中污染了食物，至少有751人。如果该小组使用了更致命的病原体，例如沙门氏菌，导致伤寒发烧，那么许多人肯定会死亡。该程序的总体目标是开发综合的等温DNA扩增和探针阵列检测幻灯片，能够快速识别各种食物和水生病原体。所有Niaid Biodefense B类食物和水源性细菌病原体大肠杆菌，纤维状霍乱，志贺氏菌痢疾，沙门氏菌物种，单核细胞增生李斯特菌，樟脑杆菌的jejuni和cookshylobacter jejuni和Enterocitia将在此程序中检测到。提出了一个能够在封闭的密封装置中实时放大和检测所有这些生物的单个集成载玻片。该计划还可以将活有机体与被食品或水卫生过程杀死的死亡生物区分开。同样，该测试可用于鉴定病原体的抗生素灵敏度，以鉴定遗传改变的生物。