Point-of-care immunoassay for early diagnosis of pertussis

用于早期诊断百日咳的即时免疫分析

• 批准号: 8780071
• 负责人: Amanda Burnham-Marusich
• 金额: $ 29.21万
• 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 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8780071
• 关键词: Antibodies; Antigen Targeting; Antigens; Bacteria; Benchmarking; Binding Sites; Bioinformatics; Biological; Biological Assay; Bordetella pertussis; Case Study; Cells; Cessation of life; Clinic; Clinical Trials; Communities; Containment; Coughing; DNA; Detection; Development; Diagnosis; Diagnostic; Disease; Disease Outbreaks; Early Diagnosis; Epitopes; Equipment; Goals; Immunoassay; Infant; International; Lateral; Life; Lung diseases; Metric; Monoclonal Antibodies; Nasopharynx; Patient Care; Patients; Pertussis; Pharyngitis; Phase; Proteins; Public Health; Resources; Respiratory Tract Infections; Sampling; Sensitivity and Specificity; Small Business Innovation Research Grant; Specificity; Staging; Streptococcal Infections; Swab; Symptoms; Target Populations; Technology; Test Result; Testing; Time; United States; Vaccines; Validation; antigen binding; base; clinically relevant; cost; design; effective therapy; improved; meetings; novel; pediatrician; phase 1 study; phase 2 study; point of care; polyclonal antibody; preclinical study; prototype; public health relevance; rapid diagnosis; research clinical testing; research study; response; success; transmission process

DESCRIPTION (provided by applicant): Pertussis is a serious and potentially life-threatening respiratory disease caused by the bacterium Bordetella pertussis. Worldwide, 18.4 million people become ill with pertussis each year, and 254,000 of these patients die. Despite high vaccine coverage in the United States, there were still over 48,000 pertussis cases reported in the U.S. in 2012. Rapid diagnosis of pertussis is critical because treatment and outbreak containment are only effective if initiated early in the disease. Currently, there are no assays that can rapidly and accurately detect early-stage pertussis. The goal is to develop a rapid, point-of-care (POC) immunoassay to detect B. pertussis during early disease. The target population will be infants who present with the non-specific, respiratory tract infection symptoms of early pertussis (prior to onset of paroxysmal cough). The approach will be detection of B. pertussis antigens from nasopharyngeal swabs by lateral flow immunoassay (LFI). The product will be an assay relevant to i) U.S. pediatrician offices, and ii) international clinics in low-resource settings: a sensitive and specific test that delivers results in 5-10min at very low cost without specialized equipment or user expertise. In preliminary studies, we used a bioinformatics analysis to identify five high priority B. pertussis protein antigens for further stdy. Specific Aim 1 will develop epitope-specific polyclonal antibodies (pAbs) for detection of B. pertussis antigens. pAbs will be screened to select those that i) react with B. pertussis proteins, ii) react with B. pertussis but not other bacteria found in the nasopharynx, iii) can function in pairs to bind the antigen in the sandwich format necessary for LFI construction, and iv) detect antigen in patient nasopharyngeal samples. Only pAbs that meet all the criteria will be advanced to LFI prototype development in Specific Aim 2. Specific Aim 2 will determine the limit of detection of B. pertussis by LFI by optimizing the physical and biological components of the LFI prototypes. Based on previously determined B. pertussis loads in infant NP swabs of 106 bacteria per swab, the benchmark for sensitivity is detection of 105 CFU per swab. Together, the experiments of Phase I will determine which B. pertussis antigen epitopes can produce pAb pairs that yield LFI prototypes with the required sensitivity. A Phase II study would develop monoclonal antibodies (mAbs) against the validated epitopes, incorporate them into an LFI, and perform detailed pre-clinical studies. Successful navigation of a full clinical trial in Phase III would ultimately yield a POC immunoassay that could dramatically increase early pertussis diagnosis, limit pertussis transmission during community outbreaks, and save patient lives.

描述（由申请人提供）：百日咳是一种由百日咳博德泰拉引起的严重且可能危及生命的呼吸道疾病。全世界每年有1840万人患百日咳，其中25.4万人死亡。尽管美国的百日咳疫苗覆盖率很高，但2012年美国报告的百日咳病例仍超过4.8万例。百日咳的快速诊断至关重要，因为只有在疾病早期开始治疗和控制疫情才有效。目前，还没有能够快速准确地检测出早期百日咳的检测方法。目标是开发一种快速的即时免疫分析方法，在疾病早期检测百日咳。目标人群将是出现非特异性呼吸道感染症状的早期百日咳（发作性咳嗽之前）的婴儿。方法将是检测百日咳抗原从鼻咽拭子侧流免疫测定（LFI）。该产品将是一种与i)美国儿科医生办公室和ii)低资源环境下的国际诊所相关的检测方法：一种敏感和特异性的检测方法，在5-10分钟内以非常低的成本提供结果，无需专业设备或用户专业知识。在初步研究中，我们使用生物信息学分析鉴定出五种高优先级百日咳蛋白抗原供进一步研究。Specific Aim 1将开发表位特异性多克隆抗体（pAbs）用于百日咳抗原的检测。将筛选pab，以选择那些i)与百日咳b杆菌蛋白反应的pab；