A Rapid Instrument Free Molecular Diagnostic for B. Pertussis

百日咳博德特氏菌的快速无仪器分子诊断

基本信息

批准号：
8718586
负责人：
Jacqueline Linnes
金额：
$ 5.91万
依托单位：
BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-02-21 至 2016-02-20
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8718586
关键词：
Acute respiratory infection Antibiotics Bacterial Infections Bedside Testings Biological Assay Blinded Blood capillaries Bordetella Bordetella pertussis Buffers Bulla Cause of Death Cessation of life Child Clinical Clinical Management Clostridium difficile Communicable Diseases Contracts DNA Detection Developing Countries Development Devices Diagnosis Diagnostic Diagnostic tests Disease Early Diagnosis Equipment Fluorescein Future Glycogen Gold Hand Hospitalization Housing Individual Infant Infection Influenza Isothiocyanates Laboratories Lateral Lung diseases Mediation Microbiology Molecular Molecular Diagnostic Testing Nucleic Acids Paper Persons Pertussis Precipitation Prevention Reaction Reagent Reproducibility Research Design Resources Respiratory Tract Infections Risk Sampling Scheme Sensitivity and Specificity Series Slide Specificity Swab Symptoms System Technology Testing Training Treatment outcome Vaccination Validation Viral Work base capillary cost experience global health health application helicase improved instrument novel point of care point-of-care diagnostics prevent public health relevance research clinical testing transmission process

项目摘要

DESCRIPTION (provided by applicant): Acute respiratory infections (ARIs) are the leading cause of death in children under five throughout the world [1]. Both vaccination and early diagnosis are critical to appropriate treatments and preventing transmission. However, infants too young to be fully immunized are most at risk of contracting ARIs. Whooping cough alone, an infection due to Bordetella pertussis (B. pertussis), causes more than 300,000 deaths per year; 90% of which occur in developing countries [2, 3]. While easily treated with first-line antibiotics more than half of all infants infected with B. pertussis require hospitalization, even in the US [2 4]. Because the initial symptoms of this bacterial infection are indistinguishable from viral ARIs, diagnosis based solely on clinical symptoms is impossible and many infants are not treated until it is too late. PCR-based molecular diagnostics performed on nasopharyngeal samples are the gold standard in B. pertussis detection. However, resource intensive equipment and technical training requirements prevent their use at the point-of-care (POC), especially in low- resource settings. A low-cost POC test for pertussis would enhance detection coverage and enable early detection resulting in improved treatment outcomes and prevention of further transmission. We hypothesize that capillary flow in ultra-low-cost paper modules can be used as a novel molecular diagnostic platform. We will develop both the individual modules for nucleic acid extraction, amplification, and detection, and their integration into a single POC device. The resulting paper-based molecular diagnostic test for B. pertussis will detect this deadly respiratory infection at the POC. The test will be hand-held, disposable, and instrument-free. This project will yield a validated proof-of-concept device suitable for future development activities including clinical testing. Ultimately, this work will develop a rapid molecular diagnosic platform that can be adapted to other diseases by simply altering the primer and probe targets in the isothermal amplification module.

描述（由申请人提供）：急性呼吸道感染（ARIS）是全世界五岁以下儿童死亡的主要原因[1]。疫苗接种和早期诊断对于适当的治疗和防止传播至关重要。但是，太年轻而无法完全免疫的婴儿最有患Aris的风险。单独咳嗽，这是由于百日咳（B. b. dustussis）引起的感染，每年造成超过300,000人死亡；其中90％发生在发展中国家[2，3]。虽然很容易用一线抗生素治疗，超过一半感染了百日咳芽孢杆菌的婴儿，即使在美国也需要住院[2 4]。因为这种细菌感染的最初症状与病毒Aris无法区分仅基于临床症状的诊断是不可能的，而且许多婴儿直到为时已晚。基于鼻咽样样品进行的基于PCR的分子诊断是百日咳芽孢杆菌检测的金标准。但是，资源密集型设备和技术培训要求阻止了他们在护理点（POC）的使用，尤其是在低资源设置中。对百日咳的低成本POC检验将增强检测覆盖范围，并使早期检测能够改善治疗结果并预防进一步传播。我们假设超低成本模块中的毛细血管流可以用作新型分子诊断平台。我们将开发核酸提取，扩增和检测的单个模块，及其整合到单个POC装置中。百日咳芽孢杆菌的基于纸张的分子诊断测试将检测到POC的这种致命的呼吸道感染。该测试将是手持式，一次性和无仪器的。该项目将产生经过验证的概念验证设备，适用于未来的开发活动，包括临床测试。最终，这项工作将开发一个快速的分子诊断平台，可以通过简单地改变等温扩增模块中的底漆和探针靶标，以适应其他疾病。