A Rapid Instrument Free Molecular Diagnostic for B. Pertussis

描述（由申请人提供）：急性呼吸道感染（阿里斯）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治疗和预防传播至关重要。然而，太小而不能完全免疫的婴儿最有可能感染阿里斯。百日咳是由百日咳杆菌（B。百日咳），每年造成超过300，000人死亡;其中90%发生在发展中国家[2，3]。虽然一线抗生素很容易治疗，但超过一半的婴儿感染了B。即使在美国，百日咳也需要住院治疗[2 4]。因为这种细菌感染的最初症状与病毒性阿里斯感染难以区分， 仅仅根据临床症状是不可能作出诊断的，许多婴儿直到为时已晚才得到治疗。对鼻咽样本进行的基于PCR的分子诊断是B的金标准。百日咳检测然而，资源密集型设备和技术培训要求阻止了它们在护理点（POC）的使用，特别是在低资源环境中。低成本的百日咳POC检测将提高检测覆盖率，并能够早期发现，从而改善治疗结果并预防进一步传播。我们假设，超低成本的纸模块中的毛细管流动可以用作一种新的分子诊断平台。我们将开发用于核酸提取、扩增和检测的单个模块，并将其集成到单个POC设备中。由此产生的B.百日咳会在POC检测到这种致命的呼吸道感染。测试将是手持式、一次性和无仪器的。该项目将产生一种经过确认的概念验证器械，适用于未来的开发活动，包括临床试验。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引物和探针靶标来适应其他疾病。

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.