A Rapid Instrument Free Molecular Diagnostic for B. Pertussis

描述(申请人提供)：急性呼吸道感染(ARI)是全世界五岁以下儿童死亡的主要原因[1]。接种疫苗和早期诊断对于适当的治疗和预防传播都至关重要。然而，太小而不能完全接种疫苗的婴儿感染ARIS的风险最大。仅百日咳这一由百日咳杆菌引起的感染每年就导致30多万人死亡，其中90%发生在发展中国家[2，3]。尽管很容易用一线抗生素治疗，但超过一半感染百日咳杆菌的婴儿需要住院治疗，即使在美国也是如此。 4]。因为这种细菌感染的最初症状与病毒ARIS难以区分， 仅根据临床症状进行诊断是不可能的，许多婴儿直到为时已晚才得到治疗。对鼻咽标本进行基于聚合酶链式反应的分子诊断是百日咳杆菌检测的金标准。然而，资源密集型设备和技术培训要求阻碍了它们在护理地点(POC)的使用，特别是在资源不足的环境中。低成本的百日咳POC检测将提高检测覆盖率，使早期检测成为可能，从而改善治疗结果并防止进一步传播。我们假设超低成本纸质组件中的毛细管流可以作为一种新型的分子诊断平台。我们将开发用于核酸提取、扩增和检测的单独模块，并将它们集成到单个POC设备中。由此产生的基于纸质的百日咳杆菌分子诊断测试将在POC检测这种致命的呼吸道感染。考试将是手持、一次性、无仪器的。该项目将产生一种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，通过简单地改变等温扩增模块中的引物和探针靶标，即可适用于其他疾病。

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.