Multiplex Screening ELISA for Giardia, Cryptosporidium and E. histolytica

贾第虫、隐孢子虫和溶组织内阿米巴多重筛选 ELISA

基本信息

批准号：
7405580
负责人：
Joel F Herbein
金额：
$ 9.95万
依托单位：
TECHLAB, INC.
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-03-15 至 2011-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7405580
关键词：
Acquired Immunodeficiency Syndrome Age All Sites Antibodies Antigens Bangladesh Biological Assay Buffers Categories Characteristics Chronic Clinical Clinical Research Collaborations Collection Communities Cost Savings Cryptosporidium Data Data Analyses Data Collection Detection Developed Countries Developing Countries Development Devices Diagnostic Diarrhea Disease Outbreaks Documentation Dose Entamoeba histolytica Enteral Environment Enzyme-Linked Immunosorbent Assay Epidemiology Evaluation Feces Food Freezing Future Germany Giardia Giardia lamblia Goals Healthcare Horseradish Peroxidase Housing Human Human Resources Human Subject Research Immunoassay In Vitro Individual Infection Intestines Japan Laboratories Manuscripts Marketing Measures Medical Surveillance Methods Microscopy Military Personnel Modification Monitor National Institute of Allergy and Infectious Disease Nature Needs Assessment Numbers Parasites Parasitic infection Pathogen detection Patients Performance Phase Phase II Clinical Trials Phase Transition Polymerase Chain Reaction Preparation Procedures Process Production Property Protocols documentation Protozoa Publications Published Comment Quality Control Recipe Records Reproducibility Research Research Ethics Committees Research Training Resolution Reverse Transcriptase Polymerase Chain Reaction Running Sampling Screening procedure Sensitivity and Specificity Shipping Ships Site Small Business Technology Transfer Research Source Specimen Standards of Weights and Measures System Testing Time Today Training United States United States Food and Drug Administration Universities Validation Variant Virginia Visit Visual Water Work base biodefense clinical research site cost cost effectiveness cross reactivity design human subject innovation pathogen prospective prototype quality assurance research and development research clinical testing response scale up size transmission process

项目摘要

DESCRIPTION (provided by applicant): Our specific objective with this fast-track STTR application is a diagnostic screen for the NIAID Biodefense Category B intestinal pathogens Giardia lamblia, Cryptosporidium spp., and Entamoeba histolytica in stool. These enteric protozoa share the characteristics of food and water-borne transmission, low infectious dose, and environmental stability. E. histolytica infections may be lethal, G. lamblia is most common, while Cryptosporidium spp. stands out as an important cause of chronic diarrhea without effective treatment in AIDS patients. The current lack of a low-cost screening fecal antigen test for enteric parasites compromises the healthcare of individuals presenting with diarrhea, as the only screening test available today is the stool O&P exam; a notoriously labor-intensive, non-specific and insensitive procedure. Our proposed diagnostic screen (the "Tri-Combo" ELISA) will provide cost-savings for clinical laboratories via the elimination from further testing specimens that are negative for all 3 parasites. The Tri-Combo ELISA is a conventional two-step ELISA format with HRP-conjugated detecting antibodies for colorimetric development. The unique multiplex nature of the test comes with its ability to simultaneously screen stool specimens for G. lamblia, Cryptosporidium spp. and E. histolytica using a single assay well. The Tri-Combo ELISA can be performed using visual interpretation of results for use in the field and in developing countries. At the same time, it is capable of conversion to automated platforms for use in reference laboratories and other large-scale surveillance scenarios; this will be useful with military and community water source outbreaks. Preliminary testing with the Tri-Combo ELISA indicates a >97% correlation to individual FDA-cleared ELISAs specific for G. lamblia, Cryptosporidium spp., and E. histolytica in stool with resolution by RT-PCR. Here, we propose optimization of the Tri-Combo ELISA in the R&D laboratory, followed by Good Manufacturing Practice (GMP) validation and FDA product clearance. Phase I will optimize the Tri-Combo ELISA, finalize GMP procedures, and prepare the clinical sites for evaluation. Phase II will complete the Tri-Combo ELISA manufacturing validation, establish performance characteristics at our consortium sites, and prepare FDA 510(k) pre-market clearance application documents. Innovation of the Tri-Combo ELISA lies with its ability to detect the three most common enteric protozoan parasites in the United States in a multiplexed format, and with its development process that allows three detection systems to be combined together in a single testing format. In the future, the process will be utilized to develop other multiplexed pathogen detection systems. A screening test is needed to detect the three most common intestinal protozoan parasites in the US (Giardia lamblia, Cryptosporidium spp., and Entamoeba histolytica). They are present in up to 10% of all stool samples submitted to reference laboratories, they are Category B Priority Biodefense Pathogens, and they share the characteristics of food and water-borne transmission, a low infectious dose, and environmental stability. Our objective is a diagnostic screen for the Giardia, Cryptosporidium, and E. histolytica in human stool specimens.

描述（由申请人提供）：我们此快速 STTR 应用的具体目标是对粪便中的 NIAID Biodefense B 类肠道病原体贾第鞭毛虫、隐孢子虫属和溶组织内阿米巴进行诊断筛查。这些肠道原虫具有食物和水传播、感染剂量低和环境稳定的特点。溶组织内阿米巴感染可能是致命的，兰伯利亚芽孢杆菌最常见，而隐孢子虫属。是艾滋病患者得不到有效治疗而导致慢性腹泻的重要原因。目前缺乏针对肠道寄生虫的低成本粪便抗原筛查，影响了腹泻患者的医疗保健，因为目前唯一可用的筛查测试是粪便 O&P 检查；这是一个众所周知的劳动密集型、非特异性和不敏感的程序。我们提出的诊断筛查（“三组合”ELISA）将通过消除对所有 3 种寄生虫均呈阴性的进一步测试样本来为临床实验室节省成本。 Tri-Combo ELISA 是一种传统的两步 ELISA 形式，具有 HRP 缀合的检测抗体用于比色显色。该测试具有独特的多重性质，能够同时筛选粪便样本中的兰氏隐孢子虫、隐孢子虫。和溶组织内阿米巴使用单个测定孔。 Tri-Combo ELISA 可以使用结果的视觉解释进行，以便在现场和发展中国家使用。同时，它能够转换为自动化平台，用于参考实验室和其他大规模监控场景；这对于军事和社区水源爆发很有用。 Tri-Combo ELISA 的初步测试表明，与经 FDA 批准的针对粪便中兰氏隐孢子虫、隐孢子虫属和溶组织内阿米巴特异性 ELISA 的相关性 > 97%，并通过 RT-PCR 进行解析。在此，我们建议在研发实验室中优化 Tri-Combo ELISA，然后进行良好生产规范 (GMP) 验证和 FDA 产品审批。第一阶段将优化 Tri-Combo ELISA、最终确定 GMP 程序并准备用于评估的临床场所。第二阶段将完成 Tri-Combo ELISA 生产验证，在我们的联盟工厂建立性能特征，并准备 FDA 510(k) 上市前许可申请文件。 Tri-Combo ELISA 的创新在于其能够以多重形式检测美国三种最常见的肠道原生动物寄生虫，并且其开发过程允许将三种检测系统以单一测试形式组合在一起。未来，该过程将用于开发其他多重病原体检测系统。需要进行筛查测试来检测美国三种最常见的肠道原生动物寄生虫（贾第鞭毛虫、隐孢子虫属和溶组织内阿米巴）。在提交给参考实验室的所有粪便样本中，它们的含量高达 10%，它们属于 B 类优先生物防御病原体，并且具有食物和水传播、低感染剂量和环境稳定性的共同特征。我们的目标是对人类粪便标本中的贾第鞭毛虫、隐孢子虫和溶组织内阿米巴进行诊断筛查。