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

• 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
• 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

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预防传播至关重要。然而，年龄太小而无法完全免疫的婴儿最有可能感染急性呼吸道感染。仅百日咳这一由百日咳博德特氏菌 (B. pertussis) 引起的感染，每年就导致超过 300,000 人死亡；其中90%发生在发展中国家[2, 3]。虽然使用一线抗生素很容易治疗，但即使在美国，超过一半的感染百日咳博德特氏菌的婴儿也需要住院治疗[2 4]。由于这种细菌感染的最初症状与病毒性 ARI 无法区分， 仅根据临床症状进行诊断是不可能的，许多婴儿等到治疗时为时已晚。对鼻咽样本进行基于 PCR 的分子诊断是百日咳博德特氏菌检测的金标准。然而，资源密集型设备和技术培训要求阻碍了它们在护理点 (POC) 的使用，特别是在资源匮乏的环境中。低成本的百日咳 POC 检测将提高检测覆盖率并实现早期检测，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模块中的毛细管流动可以用作新型分子诊断平台。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。由此产生的纸质百日咳博德特氏菌分子诊断测试将在现场检测出这种致命的呼吸道感染。该测试将是手持式、一次性且无需仪器的。该项目将产生经过验证的概念验证设备，适合未来的开发活动，包括临床测试。最终，这项工作将开发一个快速分子诊断平台，只需改变等温扩增模块中的引物和探针目标即可适应其他疾病。