Rapid multiplex method for direct phenotypic ID/AST of bacterial pathogens

用于细菌病原体直接表型 ID/AST 的快速多重方法

• 批准号: 9921292
• 负责人: Ian Fleming
• 金额: $ 116.8万
• 依托单位: GUILD ASSOCIATES, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-17 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9921292
• 关键词: Accounting; Address; Anti-Bacterial Agents; Antibiotic Resistance; Antibiotic susceptibility; Antibiotics; Antimicrobial susceptibility; Bacillus; Bacteria; Bacterial Infections; Bacteriophages; Binding; Biological Assay; Bladder Diseases; Bypass; Catheters; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Characteristics; Clinical; Cystitis; Cytolysis; DNA; Data; Detection; Development; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic tests; Disease; Engineering; Enterobacter cloacae; Escherichia coli; Female; Fluorescence; Fluorescent Probes; Genes; Genetic Engineering; Goals; Gram-Negative Bacteria; Health Care Costs; Health Personnel; Health care facility; High Prevalence; Hospitals; Human; In Vitro; Incubated; Infection; Intestines; Introns; Kidney; Kidney Diseases; Klebsiella aerogenes; Klebsiella pneumoniae; Lead; Length; Longevity; Measures; Metabolic; Methods; Microbe; Microbiology; Modernization; Patient-Focused Outcomes; Performance; Pharmaceutical Preparations; Phenotype; Predisposition; Protocols documentation; Pseudomonas aeruginosa; Public Health; RNA; RNA Splicing; Reagent; Resistance; Reverse Transcription; Ribonucleases; Septicemia; Signal Transduction; Specimen; System; Technology; Temperature; Testing; Time; Untranslated RNA; Urethra; Urinary tract infection; Urine; Virus; Visit; antimicrobial; antimicrobial drug; base; carbapenem-resistant Enterobacteriaceae; community-acquired UTI; design; diagnostic assay; extracellular; fight against; fighting; fitness; fungus; helicase; infection management; instrument; microorganism; multidrug-resistant Pseudomonas aeruginosa; nosocomial UTI; novel diagnostics; optical spectra; pathogen; pathogenic bacteria; pre-clinical; preclinical study; research and development; response; signature molecule; ureter disorder; urinary

Urinary tract infections (UTI) are diseases of the kidneys, ureters, bladder or urethra and are caused by microbes that live in the bowel. They afflict millions of people every year and are the second most common type of bacterial infection encountered by humans throughout their life span. Approximately 150 million UTI occur worldwide annually, accounting for $6 billion in healthcare costs. In the U.S.A., UTI are responsible for 8 million annual visits to healthcare providers. Some infections can lead to serious kidney complications and septicemia, with 13,000 deaths annually being attributed to nosocomial UTI. Although different microorganisms (e.g. bacteria, viruses, fungi) can cause these infections, Gram-negative bacteria are the most prevalent. The standard methods for species diagnosis are culture-based protocols that take up to 48 h. As a result, UTI are one of the most frequent reasons for antimicrobial prescriptions in healthcare facilities without a confirmed diagnosis. Therefore, modern rapid diagnostic methods that promote antimicrobial stewardship are crucial. Moreover, as patient outcomes are directly correlated to length of time to diagnosis and administration of appropriate therapy, the development of novel diagnostics that can rapidly identify (ID) the pathogen directly in clinical specimens, and simultaneously provide antibiotic susceptibility testing (AST) is a critical factor for UTI management. The goal of this project is to develop a product called multIDAST UTI that is superior to microbiological culture-based methods used for ID/AST of UTI. MultIDAST UTI will be developed as a qualitative in vitro diagnostic (IVD) test for rapid (<3 h) multiplexed identification of Gram-negative pathogens of uncomplicated UTI directly from urine and simultaneous characterization of their phenotypic responses to commonly prescribed antimicrobials. The product thereby bypasses the need for bacterial amplification and isolation and thus overcomes the major time-limiting step of current diagnostics. This will be achieved by uniquely combining species-specific phages, which have been engineered to produce a signature molecule upon bacterial infection, and isothermal helicase dependent amplification (HDA), which amplifies the signal ∼108-fold. The ensuing phenotypic signal is directly correlated to cell fitness; thus, we can rapidly generate information related to the pathogens sensitivity or resistance to a particular drug by incubation in the absence or presence of antimicrobials. Signal responses will be measured using Solana, a fluorometer currently used in multiple FDA- cleared HDA diagnostic assays. The practical purpose of this contemporary system is to identify the pathogen and determine the antibiotic suitable to cure an infection, thereby promoting antimicrobial stewardship. It will support the fight against antibiotic resistance by addressing the emergence of carbapenem-resistant Enterobacteriaceae (CRE), classified by the Centers for Disease Control as one of the nation's 'urgent' antibiotic-resistant threats.

尿路感染(UTI)是肾脏、输尿管、膀胱或尿路的疾病，由 生活在肠道中的微生物。它们每年折磨着数百万人，是第二常见的 人类一生中遇到的一种细菌感染。约1.5亿UTI 每年在全球范围内发生，占医疗成本的60亿美元。在美国，UTI负责8个 每年向医疗保健提供者提供100万人次的访问。一些感染会导致严重的肾脏并发症和 败血症，每年有13,000人死于医院内尿路感染。尽管不同的微生物 (如细菌、病毒、真菌)可引起这些感染，其中以革兰氏阴性菌最为普遍。这个 物种诊断的标准方法是基于培养的方法，需要长达48小时。因此，UTI 在医疗机构未经确认的情况下开出抗菌药物处方的最常见原因之一 诊断。因此，促进抗菌药物管理的现代快速诊断方法至关重要。 此外，由于患者的预后与诊断和给药的时间长短直接相关 适当的治疗，开发新的诊断方法，可以直接快速识别(ID)病原体 临床标本，同时提供药敏试验(AST)是尿路感染的关键因素 管理层。 这个项目的目标是开发一种名为MULTIDAST UTI的产品，它优于微生物 UTI的ID/AST采用基于文化的方法。MULTIDAST UTI将在体外作为定性研究开发 诊断(IVD)试验快速(&lt；3；h)多重鉴定非合并症革兰阴性杆菌 直接从尿液中分离的UTI及其对常见疾病的同时表型反应 开出的抗菌药。因此，该产品省去了细菌扩增和分离的需要， 从而克服了当前诊断的主要时间限制步骤。这将通过独特的组合来实现 物种特异性噬菌体，已经被设计成在细菌上产生一个标志性的分子 感染，以及等温解旋酶依赖的扩增，将信号∼放大108倍。这个 随后的表型信号与细胞适合度直接相关；因此，我们可以快速生成相关信息 对病原体的敏感性或抗药性，在没有或存在的情况下孵育 抗菌剂。信号响应将使用Solana来测量，Solana是一种目前在多个FDA- 已通过HDA诊断化验。这一现代系统的实际目的是识别病原体。 并确定适合治愈感染的抗生素，从而促进抗微生物管理。会的 通过解决碳青霉烯类耐药菌的出现来支持对抗抗生素耐药性的斗争 肠杆菌科(CRE)，被疾病控制中心列为国家紧急事件之一 抗生素抗药性威胁。