Smartphone-linked system for diagnosis and epidemiological reporting of pathogens at the point of care

智能手机连接系统，用于在护理点诊断和流行病学报告病原体

基本信息

批准号：
10462690
负责人：
Rashid Bashir
金额：
$ 40.48万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-05 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10462690
关键词：
Algorithms Animals Bar Codes Base Sequence Biological Assay Blood Blood specimen Bluetooth Brazil Buffers Capital Cellular Phone Cessation of life Chemicals Clinic Clinical Communicable Diseases Communication Complex Cytolysis DNA-Directed DNA Polymerase Data Databases Dengue Deposition Detection Development Diagnosis Diagnostic tests Differential Diagnosis Disease Notification Environmental Monitoring Epidemiology Equipment Fingers Flavivirus Fluorescence Food Safety Geography Goals Gold Health Health Food Health Services Hospitals Human Image Image Analysis Infection Insect Vectors Institutes Institution International Kinetics Laboratories Lighting Link Liquid substance Measurement Mediating Microfluidic Microchips Microfluidics Microprocessor Monitor Nucleic Acids Optics Outcome Pathogen detection Patients Physicians Plasmids Polymerase Chain Reaction Population Process Protocols documentation Public Health Reaction Reagent Reporting Resolution Resources Reverse Transcription Sampling Secure Serum Services Site Specificity Students Symptoms System Technology Testing Time Translating Viral Whole Blood World Health Organization ZIKA Zika Virus assay development authority base biochip burden of illness chikungunya chromatin immunoprecipitation cloud based communicable disease diagnosis cost design detection limit detection platform digital disability epidemiologic data fluorescence imaging health management image processing imaging modality instrument interest isothermal amplification mortality multiplex detection novel novel strategies pathogen pathogenic bacteria pathogenic virus point of care point of care testing portability public health emergency repository sensor service organization smartphone Application tool wireless

项目摘要

Abstract Infectious disease remains the world’s top contributor to death and disability, and, due to their ability to spread rapidly through insect vectors or contact with bodily fluids, there is an urgent need for simple, sensitive and easily translatable point-of-care tests, particularly for infections that are difficult to differentiate based upon clinical symptoms alone. This project develops a novel point-of-care platform to quantitatively diagnose viral infectious diseases (Zika, Dengue (types 1 and 3), and Chikungunya) from whole blood samples using a microfluidic platform that performs sample pre-processing in a microfluidic cartridge followed by real-time reverse-transcription loop-mediated isothermal amplification (RT-LAMP) in the same cartridge with pre-dried primers specific to pathogen targets. Our handheld, inexpensive (~$500), point-of-care platform communicates its measurements to a smartphone to process a sequence of fluorescence images of the amplification reaction. Image analysis of the dynamic amplification process is used to estimate the pathogen count from amplification initiation points using a novel approach called “spatial” LAMP (S-LAMP). Our preliminary data shows that the approach provides selectivity and detection limits that are equivalent to conventional Polymerase Chain Reaction (PCR) or LAMP reactions performed with conventional laboratory instruments, and the ability to detect the targets with high specificity from complex media. The S-LAMP approach demonstrates potential to detect 1-5 pathogen copies per reaction. The system will be evaluated using a rigorous tiered approach using plasmids containing the target nucleic acid sequence for initial characterization of detection limits, followed by nucleic acid pathogen extracts, culminating of detection of the pathogens in whole blood using chemical lysis to release target nucleic acids. Partnering with collaborators at the Institute for Infectious Diseases in Brazil, and Carle Clinic in Urbana, the system will be utilized on patient sample repositories and measurements compared to gold standard analysis. Importantly, as detection, image processing, and quantitation are performed with the smartphone microprocessor, detection is easily integrated with a cloud-based database reporting system that facilitates communication with remotely-located physicians and tracking of epidemiological data by health services. The resulting platform will be broadly applicable to a wide variety of multiplexed panels of pathogens that are of interest for human health, animal health, food safety, and environmental monitoring.

抽象的传染病仍然是世界上导致死亡和残疾的首要原因，并且由于其传播能力通过昆虫媒介或与体液接触迅速传播，迫切需要简单、敏感和易于翻译的即时检测，特别是对于难以区分的感染该项目开发了一种新颖的护理点平台来定量诊断病毒。使用全血样本检测传染病（寨卡病毒、登革热（1 型和 3 型）和基孔肯雅热）微流控平台，在微流控盒中执行样品预处理，然后进行实时处理在同一盒中进行逆转录环介导等温扩增 (RT-LAMP) 并预干燥我们的手持式廉价（约 500 美元）护理点平台可进行通信。它对智能手机进行测量，以处理扩增反应的一系列荧光图像。动态扩增过程的图像分析用于估计扩增中的病原体计数我们的初步数据表明，使用一种称为“空间”LAMP（S-LAMP）的新方法的起始点。方法提供与传统聚合酶链相当的选择性和检测限使用传统实验室仪器进行的反应 (PCR) 或 LAMP 反应，以及能够 S-LAMP 方法具有从复杂介质中检测高特异性目标的潜力。每个反应检测 1-5 个病原体拷贝该系统将使用严格的分层方法进行评估。含有目标核酸序列的质粒用于检测限的初始表征，然后核酸病原体提取物，最终使用化学裂解检测全血中的病原体与巴西传染病研究所的合作者合作释放目标核酸，和厄巴纳的卡尔诊所，该系统将用于患者样本存储库和测量重要的是，与金标准分析相比，检测、图像处理和定量都是如此。通过智能手机微处理器执行检测，可轻松与基于云的数据库集成报告系统，促进与远程医生的沟通和跟踪卫生服务部门的流行病学数据将广泛适用于各种领域。对人类健康、动物健康、食品安全和健康感兴趣的多重病原体组环境监测。