Portable magnetofluidic platform for rapid multiplexed detection of sexually transmitted infections

用于快速多重检测性传播感染的便携式磁流体平台

• 批准号: 10546170
• 负责人: Alexander Trick
• 金额: $ 25.86万
• 依托单位: PROMPT DIAGNOSTICS LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-10 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10546170
• 关键词: Address; Adoption; Algorithms; Antibiotic Therapy; Antimicrobial Resistance; Automation; Azithromycin; Biological Assay; Buffers; Categories; Cefixime; Ceftriaxone; Cephalosporins; Chlamydia Infections; Chlamydia trachomatis; Ciprofloxacin; Clinic; Clinical; Code; Color; Complex; Contact Tracing; Contracts; Coupled; Critical Care; Custom; DNA; DNA Probes; Data; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Dyes; Engineering; Epidemic; Epidemiology; Fluorescence; Foundations; Funding; Genetic Markers; Genotype; Gonorrhea; HIV; Heating; Human; Image; Individual; Infection; Life; Link; Liquid substance; Magnetism; Morbidity - disease rate; Neisseria gonorrhoeae; Pathogenicity; Patients; Performance; Phase; Precision therapeutics; Predisposition; Public Health; Ramp; Reaction; Reagent; Resistance; Resistance development; Resistance profile; Resistance to infection; Resolution; Resource-limited setting; Risk; Running; Sampling; Sensitivity and Specificity; Sexually Transmitted Diseases; Shoes; Signal Transduction; Small Business Innovation Research Grant; Specificity; Spectinomycin; Syndrome; Syphilis; Techniques; Temperature; Test Result; Testing; Time; Trichomonas Infections; Trichomonas vaginalis; United States; Validation; Work; World Health Organization; antimicrobial; antimicrobial drug; assay development; base; clinically relevant; combat; cost; design; detection assay; detection limit; detection platform; diagnostic platform; disability; emerging antimicrobial resistance; fluorophore; innovation; instrument; instrumentation; magnetic beads; melting; miniaturize; mortality; multiplex assay; multiplex detection; operation; optimal treatments; overtreatment; pathogen; point of care; point of care testing; portability; reference genome; resistance gene; response; screening; transmission process; two-dimensional

Project Summary/Abstract The 21st century has seen a rise in sexually transmitted infections (STIs) to epidemic proportions with recent estimates of over 1 million curable STIs acquired daily. To combat the spread of STIs, diagnostics that are fast, sensitive, and affordable are needed to accurately identify pathogens and link patients to the appropriate treatment. Antimicrobial resistance (AMR), particularly in Neisseria gonorrhoeae (NG), threatens to aggravate the spread of STIs with untreatable infections, so tests that also characterize the AMR profile of infections are needed to identify the proper treatment and promote antimicrobial stewardship. Currently available diagnostics are either too slow to guide prescribed treatment, too expensive for widespread adoption, or too limited to provide a complete diagnosis. In this Phase I SBIR proposal, we will develop a 12-plex assay for detection of the 3 most prevalent STI pathogens- NG, Chlamydia trachomatis (CT), Trichomonas vaginalis (TV)- along with genotypic characterization of NG AMR targets for 4 types of antimicrobials. This assay will be integrated into low-cost magnetofluidic cartridges for fully automated sample-to-answer operation in a portable instrument for use at the point-of-care. Magnetofluidic cartridges leverage magnetic transfer of functionalized beads through preloaded reagents to capture, purify, and transport analytes without the need for complex fluidic circuitry and controls that increase cost of diagnostic platforms currently on the market. Our simple extruded thermoplastic well design enables easily scalable manufacture and focused heating using a miniaturized heat block for rapid PCR with turnaround times under 25 minutes. High degrees of multiplexing will be achieved in part using our recent developments in two-dimensional/two-dye (2D)-melt assays which identify amplified PCR targets using probes encoded with both fluorophore dye color and hybridization melt temperature. Another key innovation for multiplexing within our cartridges augments our previously demonstrated magnetofluidic capabilities to enable distribution of captured DNA between multiple reaction wells via a sequential multi-elution technique. The proposed work in this Phase I project will lay the foundation for a clinically relevant test with 2D- melt assay development (Aim 1), expansion of our Prompt magnetofluidic platform to run the 2D-melt assays (Aim 2), and validation of our assays on reference genomes and clinical isolates (Aim 3). Subsequent Phase II funding will allow design refinement for manufacturing cartridges with automated assembly, prolonged shelf-life, and human factors engineering.

项目总结/摘要 21世纪世纪，性传播感染（STIs）上升到流行病的比例， 估计每天有100多万例可治愈的性传播感染。为了防止性传播感染的传播，快速诊断， 我们需要灵敏、经济的设备来准确识别病原体， 治疗抗生素耐药性（AMR），特别是淋病奈瑟菌（NG），有可能加剧 无法治疗的感染性传播疾病的传播，因此也表征感染的AMR特征的测试是 需要确定适当的治疗方法并促进抗菌药物管理。当前可用的诊断程序 要么太慢，无法指导规定的治疗，要么太昂贵，无法广泛采用，要么太有限，无法提供 完整的诊断。在这一I期SBIR提案中，我们将开发一种12重检测方法，用于检测3种 最流行的STI病原体- NG、沙眼衣原体（CT）、阴道毛滴虫（TV）-沿着 4种类型抗菌剂的NG AMR靶标的基因型表征。本检测试剂盒将整合 低成本的磁流体盒，用于便携式设备中的全自动采样-应答操作， 在护理点使用的仪器。磁流体盒利用功能化的磁性转移 珠通过预载的试剂来捕获、纯化和运输分析物，而不需要复杂的流体 增加了目前市场上诊断平台的成本。我们简单的挤出 热塑性井设计使得能够容易地可扩展制造和使用小型化加热器集中加热 用于快速PCR的阻断，周转时间低于25分钟。高度的多路复用将在 部分使用我们最近在二维/双染料（2D）-熔解分析中的发展， 使用用荧光团染料颜色和杂交解链温度编码的探针靶向。另一个关键 在我们的盒内进行多路复用的创新增强了我们先前展示的磁流体 能够通过连续的多次洗脱在多个反应威尔斯孔之间分配捕获的DNA 法该第一阶段项目中的拟议工作将为2D临床相关测试奠定基础- 熔解试验开发（目标1），扩展我们的Prompt磁流体平台以运行2D熔解试验 (Aim 2），并验证我们的检测参考基因组和临床分离株（目的3）。后续第二阶段 资金将允许设计改进，用于制造自动组装，延长保质期， 和人为因素工程。