Detection of Biodefense Pathogens Using Phage Diagnostics

使用噬菌体诊断检测生物防御病原体

• 批准号: 8819100
• 负责人: DAVID A SCHOFIELD
• 金额: $ 74.54万
• 依托单位: GUILD ASSOCIATES, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-07 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8819100
• 关键词: Address; Aldehydes; Anthrax disease; Antibiotic susceptibility; Antibiotics; Antimicrobial susceptibility; Automation; Bacillus anthracis; Bacteriophages; Binding; Biological Assay; Biological Warfare; Categories; Cell Separation; Cells; Centers for Disease Control and Prevention (U.S.); Characteristics; Chromosomes; Clinical; Collection; Cytolysis; Data; Data Analyses; Detection; Development; Devices; Diagnosis; Diagnostic; Diagnostic Specificity; Disease; Disease Outbreaks; Disease Progression; Early Diagnosis; Engineering; Ensure; Enzymes; FDA approved; Freeze Drying; Genes; Genome; Goals; Growth; Health; Infection; Laboratories; Light; Luciferases; Lytic; Measures; Medical; Methodology; Methods; National Institute of Allergy and Infectious Disease; Onset of illness; Outcome; Patients; Performance; Phenotype; Plague; Process; Production; Public Health; Reagent; Recombinants; Recording of previous events; Reporter; Reporter Genes; Research; Signal Transduction; Specificity; Specimen; Staging; Strategic Planning; Symptoms; Technology; Testing; Time; Translating; Yersinia pestis; absorption; antimicrobial; antimicrobial drug; base; biodefense; clinically relevant; cost; cost effective; diagnostic assay; disorder prevention; fighting; improved; mortality; outcome forecast; pathogen; pre-clinical; product development; prototype; rapid detection; rapid diagnosis; reagent testing; receptor; scale up; screening; tool

DESCRIPTION (provided by applicant): Yersinia pestis and Bacillus anthracis are designated by the Centers for Disease Control and Prevention (CDC) as Category A bacterial pathogens and are the etiological agents for the plague and anthrax, respectively. Although the natural occurrence of each disease is rare, the possibility of terrorist groups using these pathogens as bioweapons is real. Because both conditions share common features such as rapid clinical course, and high mortality, it is critical that outbreaks be detected quickly. Therefore, methodologies that provide rapid detection and diagnosis are essential to ensure immediate implementation of public health measures and activation of crisis management. The goal of this research is to develop a diagnostic platform for the plague and anthrax. The CDC currently uses classical phage lysis assays as a standard for the confirmed identification of Y. pestis and B. anthracis. Although these assays are robust, they are laboratory based and require 24-48 h to complete. In order to overcome these limitations, we have generated recombinant "light-tagged"-reporter phages which can rapidly (within minutes), sensitively and specifically transduce a bioluminescent phenotype to Y. pestis and B. anthracis. We have demonstrated that the reporter phages can detect the target pathogens at clinically relevant concentrations during the early course of disease, and also generate antibiotic susceptibility profiles 5 to 10 times faster than conventional methods. This is key as the diseases are treatable as long as they are rapidly diagnosed, and appropriate therapy is initiated. We will continue with this research and transition the diagnostic technology to preclinical development with the goal of gaining FDA-approval. In the "NIAID: Strategic Planning for the 21st Century", NIAID highlights the following priorities in the development of diagnostic assays: (i) specific to the pathogen; (ii cost effective; (iii) sensitive; (iv) multiplexed and high throughput, and (v) easy to use. This proposal is significant because the reporter phage technology can address these requirements, and has the potential to save lives by significantly improving our nation's defense against biological warfare.

描述（由申请人提供）：鼠疫耶尔森菌和炭疽芽孢杆菌被美国疾病控制和预防中心（CDC）指定为A类细菌病原体，分别是鼠疫和炭疽的病原体。虽然每种疾病的自然发生都很罕见，但恐怖主义团体利用这些病原体作为生物武器的可能性是真实的。由于这两种疾病具有共同的特征，如快速的临床过程和高死亡率，因此迅速发现疫情至关重要。因此，提供快速检测和诊断的方法对于确保立即执行公共卫生措施和启动危机管理至关重要。这项研究的目标是开发一个鼠疫和炭疽的诊断平台。CDC目前使用经典的噬菌体裂解试验作为确认鉴定Y的标准。鼠疫和B.炭疽病虽然这些测定是稳健的，但它们是基于实验室的，需要24-48小时才能完成。为了克服这些局限性，我们已经产生了重组的“光标记”-报告基因，它可以快速（在几分钟内），敏感和特异性地将生物发光表型标记为Y。鼠疫和B.炭疽病我们已经证明，报告基因可以在疾病的早期过程中检测到临床相关浓度的目标病原体，并且产生抗生素敏感性谱的速度比传统方法快5至10倍。这一点很关键，因为只要迅速诊断出这些疾病，并开始适当的治疗，这些疾病是可以治疗的。我们将继续这项研究，并将诊断技术过渡到临床前开发，目标是获得FDA批准。 在“NIAID：NIAID在“21世纪世纪战略规划”中强调了以下诊断检测开发的优先事项：（i）病原体特异性;（ii）成本效益;（iii）敏感性;（iv）多重和高通量，以及（v）易于使用。这一提议意义重大，因为报告噬菌体技术可以满足这些要求，并有可能通过显着提高我们国家对生物战的防御来拯救生命。