Point-of-care lateral flow immunoassay for diagnosis of pertussis

用于诊断百日咳的即时侧流免疫分析

• 批准号: 10693572
• 负责人: Amanda Burnham-Marusich
• 金额: $ 100.94万
• 依托单位: DXDISCOVERY, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10693572
• 关键词: Affinity; Agreement; Annual Reports; Antibodies; Antigen Targeting; Antigens; Bacteria; Blood group antigen S; Bordetella pertussis; COVID-19; COVID-19 pandemic; Case Study; Categories; Cell surface; Cells; Clinic; Clinical; Commercial grade; Communities; Coughing; Country; Data; Detection; Diagnosis; Diagnostic; Disease; Disease Outbreaks; Early Diagnosis; Early treatment; Electricity; Epitope Mapping; Equipment; Goals; Healthcare; Hospitals; Hour; Immunoassay; Infant; Infection; Laboratories; Lateral; Libraries; Linear Sequence Epitopes; Measures; Modeling; Monoclonal Antibodies; Morbidity - disease rate; Nasopharynx; Nucleic Acids; Papio; Patients; Performance; Pertussis; Phase; Preparation; Proteins; Protocols documentation; Public Health; Rapid diagnostics; Reaction; Reproducibility; Research; Resources; Respiratory Disease; Sampling; Sensitivity and Specificity; Serology; Severity of illness; Specificity; Specimen; Speed; Statistical Data Interpretation; Symptoms; Target Populations; Testing; Trachea; United States; Vaccines; Visual; antigen detection; antigen test; clinically relevant; community transmission; coronavirus disease; cross reactivity; design; detection limit; detection sensitivity; economic impact; factor A; fungus; illness length; improved; microbial; nasopharyngeal swab; pediatrician; point of care; point of care testing; pre-clinical; preclinical evaluation; prevent; prospective; prototype; rapid test; research clinical testing; specific biomarkers; tool; transmission process; urgent care; waiver

Pertussis (Whooping cough) is a highly contagious, airborne disease that is rapidly re-emerging as a serious public health threat in the United States. Despite high vaccine coverage, annual reported cases in the U.S. have been increasing recently, with an average of 25,000 per year from 2010-2019. This is a level unseen since the early 1960s. Diagnosis of pertussis is challenging because its early symptoms (during the contagious catarrhal stage, prior to paroxysmal cough) overlap with other respiratory diseases. This is analogous to the diagnostic challenge of COVID-19. The ongoing COVID-19 pandemic has illustrated the critical public health importance of accessible, rapid diagnostics for controlling the spread of highly transmissible respiratory diseases, a category that includes pertussis. Consequently, as with COVID-19, there is a critical need to develop improved pertussis diagnostics to enable prompt, appropriate patient treatment as well as limit needless community transmission. The project goal is a rapid, point-of-care (POC), lateral flow immunoassay (LFIA) to detect Bordetella pertussis antigen from nasopharyngeal specimens. The target population will be symptomatic patients who present in pediatrician offices, urgent care clinics, or small hospital ERs. The product will be a sensitive and specific test that enables immediate action, akin to other highly used and accepted LFIAs (e.g. COVID-19, RSV). This proposal builds on data and results from Phases I and II. First, the B. pertussis protein tracheal colonization factor A (TcfA) was identified as a specific biomarker for presence of the bacterium. Second, a library of epitope- mapped monoclonal antibodies (mAbs) with high affinity and specificity to TcfA was developed. Third, over 784 permutations of mAb combinations were evaluated for construction of a LFIA. Fourth, the prototype LFIA’s configuration was optimized for high specificity and sensitivity for detection of both purified TcfA and clinically relevant concentrations of B. pertussis cells. Fifth, the prototype LFIA showed high sensitivity and specificity for diagnosis of pertussis in a baboon model. Finally, proof-of-concept was demonstrated with patient specimens. Per the original plan, Phase IIb builds on Phases I and II to deliver an FDA-cleared diagnostic. Three aims are proposed. Aim 1 includes an FDA Pre-Submission and transitions from a research-grade, prototype LFIA to a commercial-grade product made under GMP. Aim 2 is a pre-clinical evaluation of precision and reproducibility, limit of detection, analytical specificity and inclusivity, stability, ruggedness, interfering substances, and microbial interference. Aim 3 is a clinical evaluation that will end with a FDA 510(k) Submission. If successful, this study will deliver a pertussis POC test that is easily accessible to all clinics and that provides results in an actionable timeframe – thereby reducing patient morbidity and preventing unnecessary community transmission.

百日咳（百日咳）是一种高度传染性的空中疾病，正迅速重新出现为严重 美国的公共卫生威胁。尽管疫苗覆盖范围很高，但美国的年度报告案件 最近有所增加，从2010年至2019年，平均每年25,000。这是一个看不见的水平，因为 1960年代初。百日咳的诊断是具有挑战性的，因为它的早期症状（在传染性的卡塔哈尔期间 阶段，在阵发性咳嗽之前）与其他呼吸道疾病重叠。这类似于诊断 COVID-19的挑战。持续的共同19-19大流行已经说明了公共健康的关键重要性 可访问的快速诊断，用于控制高度可传播的呼吸疾病的传播，这是一个类别 包括百日咳。因此，与COVID-19一样，迫切需要改善的百日咳 诊断能够迅速，适当的患者治疗以及限制不必要的社区传播。 项目目标是快速的，即时的（POC），侧向流免疫测定（LFIA），以检测百日咳bordetella belttussis 鼻咽标本的抗原。目标人群将是有症状的患者 儿科医生办公室，紧急护理诊所或小型医院。该产品将是一个敏感且特定的测试 这样可以立即采取行动，类似于其他高度使用和接受的LFIA（例如Covid-19，RSV）。 该建议基于数据和II阶段的结果。首先，百日咳蛋白质气管定植 因子A（TCFA）被确定为存在细菌的特定生物标志物。第二，一个表位库 - 开发了具有高亲和力和特异性TCFA的映射的单克隆抗体（mAb）。第三，超过784 评估了MAB组合的排列以构建LFIA。第四，原型LFIA 优化配置以高特异性和敏感性，以检测纯化的TCFA和临床 百日咳细胞的相关浓度。第五，原型LFIA显示出高灵敏度和特异性 狒狒模型中百日咳的诊断。最后，用患者标本证明了概念验证。 根据原始计划，IIB期建立在I和II阶段，以提供FDA清除诊断。三个目标是 建议的。 AIM 1包括FDA预提取和从研究级原型LFIA到A的过渡 根据GMP生产的商业级产品。 AIM 2是对精度和可重复性的临床前评估， 检测极限，分析特异性和包容性，稳定性，坚固性，干扰物质和微生物 干涉。 AIM 3是临床评估，将以FDA 510（k）提交结束。如果成功，这项研究 将提供一项百日咳POC测试，所有诊所都很容易获得，这为可行的结果提供了结果 时间范围 - 从而降低患者的发病率并防止不必要的社区传播。