Technology development for point-of-care detection and antimicrobial susceptibility testing of Neisseria gonorrhoeae

淋病奈瑟菌即时检测和药敏试验技术开发

• 批准号: 9768322
• 负责人: CHARLOTTE Ann GAYDOS
• 金额: $ 126.72万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-22 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9768322
• 关键词: Address; Adopted; Antimicrobial Resistance; Antimicrobial susceptibility; Azithromycin; Bacteria; Bacterial DNA; Bacterial RNA; Baltimore; Biological Assay; Ceftriaxone; Centers for Disease Control and Prevention (U.S.); Chemistry; Ciprofloxacin; Cities; Clinic; Clinical; Clinical Management; Country; DNA Markers; Data; Detection; Development Plans; Device or Instrument Development; Devices; Diagnosis; Diagnostic; Drug Controls; Drug resistance; Evaluation; Evolution; Freeze Drying; Future; Genes; Goals; Gonorrhea; Guidelines; Health; Hour; Individual; Industrialization; Infection; Measures; Membrane; Methods; Microbiology; Microfluidics; Mineral Oil; Molecular; Neisseria gonorrhoeae; Nucleic Acid Amplification Tests; Nucleic Acids; Oils; Optics; Patients; Pharmaceutical Preparations; Phenotype; Physiological; Predisposition; Protocols documentation; Pseudogenes; Publishing; Reaction; Reagent; Reporting; Resistance; Resistance development; Resistance profile; Reverse Transcriptase Polymerase Chain Reaction; Reverse Transcription; Risk; Role; Sampling; Scheme; Speed; Swab; System; Systems Integration; Technology; Testing; Time; Translations; Vacuum; Validation; antimicrobial; base; cell growth; design; drug testing; innovation; instrument; laboratory facility; molecular diagnostics; pathogen; personalized medicine; point of care; portability; product development; programs; rRNA Precursor; reconstitution; resistant strain; sample collection; success; technology development; treatment guidelines; validation studies

PROJECT SUMMARY Antimicrobial resistance (AMR) in Neisseria gonorrhoeae (NG) is in the top tier of AMR l threats as defined by WHO. Over the past decades, NG has developed resistance to all antimicrobials previously recommended for treatment of gonorrhea, leaving dual therapy of ceftriaxone plus azithromycin as currently the only appropriate option for empirical first-line therapy in most countries world-wide. Now NG strains have been reported resistant to both ceftriaxone and azithromycin. Conversely, although ciprofloxacin is no longer recommended by the CDC for the treatment of NG, recent studies suggest that a large percentage of GC infections could be potentially treated with ciprofloxacin. With the continued evolution of AMR, there is an urgent need for personalized treatment approaches that target an individual infection, in contrast to the current “globally uniform” empiric approach. However, this requires clinicians to know drug resistance or susceptibility quickly enough to inform prescription decisions. To mitigate the emergence and spread of AMR in NG, CDC periodically publishes STD treatment guidelines to assist clinicians. These guidelines are informed by susceptibility data generated by the national CDC Gonococcal Isolate Surveillance Project (GISP). GISP’s impact, however, is being jeopardized by technological evolution. Determining AMR requires prolonged (24-48 hours) microbiological cultivation in sophisticated laboratory facilities. But the advent of nucleic acid amplification tests (NAAT) with enhanced speed and accuracy has supplanted culture-based diagnosis of NG infections, leading to limited specimen collection for culture and loss of capability to perform culture of NG in most testing clinics. Consequently, the success of widespread NAAT has inadvertently created a critical void in AMR testing. We propose to develop a complete diagnostic solution capable of performing identification (ID) of NG infection and phenotypic antimicrobial susceptibility testing (AST). Specifically, ID is achieved using PCR to detect NG- specific DNA markers; while AST is carried out using quantitative PCR to measure the difference in nucleic acid markers (bacterial DNA or RNA) which correlate with the physiologic state of pathogen between drug- treated samples and no-drug controls. Our combined ID-AST platform, which capitalizes on innovative advances in NAAT and microfluidics, has the potential to deliver all essential NG diagnostic information specific to each suspected patient at the POC to tailor personalized treatment; its practical design is also well- suited to resolve the technical challenges confronting GISP for routine surveillance of NG AMR. We propose the following aims: 1) to develop a streamlined diagnostic protocol for integrated ID and AST of NG; 2) to develop a droplet microfluidic cartridge implementing the integrated ID-AST assay; 3) system integration and instrument development; and 4) analytical and clinical validation of the integrated system. To facilitate technology translation, a Product Development Plan for future clinical deployment is proposed.

项目概要 淋病奈瑟菌 (NG) 的抗菌素耐药性 (AMR) 属于 AMR l 威胁的顶级，定义如下： 在过去的几十年里，NG 已经对以前推荐的所有抗菌药物产生了耐药性。 治疗淋病，头孢曲松加阿奇霉素的双重治疗是目前唯一合适的 目前，世界上大多数国家都已报道了 NG 菌株的经验性一线治疗。 尽管不再推荐环丙沙星，但对头孢曲松和阿奇霉素均具有耐药性。 CDC 治疗 NG 的方法，最近的研究表明，很大一部分 GC 感染可能是由 随着 AMR 的不断发展，迫切需要环丙沙星来治疗。 与当前的“全球范围内的治疗方法”相比，针对个体感染的个性化治疗方法 然而，这需要各团快速了解耐药性或敏感性。 足以为处方决策提供信息。 为了减少 NG 中 AMR 的出现和传播，CDC 定期发布 STD 治疗指南 这些指南是根据国家疾病预防控制中心生成的敏感性数据提供的。 然而，淋球菌分离监测项目 (GISP) 的影响正受到技术的威胁​​。 确定 AMR 需要在复杂的环境中进行长时间（24-48 小时）的微生物培养。 但核酸扩增测试 (NAAT) 的出现提高了速度和效率。 NG 感染的准确性已经取代了基于培养的诊断，导致 NG 感染的标本采集有限 在大多数检测诊所中，培养和丧失进行 NG 培养的能力，是成功的。 广泛的 NAAT 无意中在 AMR 测试中造成了严重的空白。 我们建议开发一套能够执行 NG 感染识别 (ID) 的完整诊断解决方案 具体来说，ID是通过PCR检测NG-来实现的。 特异性DNA标记；而AST则使用定量PCR来测量核酸的差异 与药物之间病原体的生理状态相关的酸标记（细菌DNA或RNA） 我们的组合 ID-AST 平台利用了创新技术。 NAAT 和微流体技术的进步，有潜力提供所有重要的 NG 诊断信息 针对POC的每一位疑似患者量身定制个性化治疗方案，其实用性设计也很好—— 适合解决 GISP 日常监测 NG AMR 所面临的技术挑战。 目标如下： 1) 开发 NG 的集成 ID 和 AST 的简化诊断方案 2) 开发实现集成 ID-AST 的液滴微流体盒 3) 系统集成和检测； 仪器开发；4) 集成系统的分析和临床验证。 技术转化，提出了未来临床部署的产品开发计划。