Synthetic Peptide SARS Coronavirus Diagnostic Kit

合成肽SARS冠状病毒诊断试剂盒

• 批准号: 6920669
• 负责人: Chang Y. Wang
• 金额: $ 76.27万
• 依托单位: UNITED BIOMEDICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-15 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6920669
• 关键词: China; SARS virus; Taiwan; United States; biomedical automation; clinical trials; communicable disease diagnosis; cooperative study; diagnosis design /evaluation; diagnostic tests; emerging infectious disease; enzyme linked immunosorbent assay; human tissue; patient oriented research; peptide chemical synthesis; serology /serodiagnosis; severe acute respiratory syndrome; virus antigen; virus genetics; virus protein

DESCRIPTION (provided by applicant): The long-term objectives are to develop an enzyme-linked immunoassay (ELISA) for Severe Acute Respiratory Syndrome (SARS), based on synthetic peptide antigens. The test will be stable and free of biohazardous manufacturing operations and components, and is intended as a safe and rapid method for serological surveillance. The test will be amenable to large scale manufacture and compatible with fully automated diagnostic screening. Specific aims are 1) to identify and optimize the peptide antigens, including antigens from non-structural SARS coronavirus (SCoV) proteins, in addition to peptides from structural proteins S, M, and N that have already been identified; 2) formulate a SARS ELISA test kit optimized for sensitivity and specificity; 3) validate the kit with a panel of characterized sera from SARS patients and various panels from the sera of subjects that have not been exposed to the SCoV; 4) perform a large-scale clinical trial at multiple sites in the US, Taiwan and China; 4) accomplish the commercial scale production of kits; and 5) submit a PMA or PDP application for US and foreign regulatory approvals. A prototype SARS ELISA having peptide antigens from the S, M, and N structural proteins of SCoV is to be improved by the addition of non-structural peptides whose sequences are deduced from the putative pol region of the SCoV genome. Overlapping peptides will be synthesized to cover the entire pol la and lb open reading frames. Peptides will be of lengths from 20-60 amino acids. Individual peptides will be screened by the ELISA method for reactivity to a collection of sera from confirmed SARS patients. Serologically reactive epitopes on the peptides will be refined for optimum sequences and conformation. The newly identified non-structural peptide antigens will be added to the solid-phase antigen formulation of the SARS ELISA. The peptides and kits will be manufactured and characterized with cGMP standards. Large-scale clinical trials will be done at facilities in Asia on extensive collections of SARS positive sera.

描述（由申请人提供）： 长期目标是开发一种基于合成肽抗原的用于严重急性呼吸综合征（SARS）的酶联免疫测定（ELISA）。该测试将稳定且不含生物危害的制造操作和成分，旨在作为一种安全、快速的血清学监测方法。该测试将适合大规模生产，并与全自动诊断筛查兼容。具体目标是1）鉴定和优化肽抗原，除了已经鉴定的结构蛋白S、M和N的肽之外，还包括非结构SARS冠状病毒（SCoV）蛋白的抗原； 2) 制定敏感性和特异性优化的SARS ELISA检测试剂盒； 3) 使用来自 SARS 患者的一组特征血清和来自未接触过 SCoV 的受试者血清的各种组来验证该试剂盒； 4）在美国、台湾和中国大陆多个地点进行大规模临床试验； 4）完成试剂盒的商业化规模生产； 5) 提交 PMA 或 PDP 申请以获得美国和外国监管机构的批准。具有来自 SCoV 的 S、M 和 N 结构蛋白的肽抗原的 SARS ELISA 原型将通过添加非结构肽进行改进，这些非结构肽的序列是从 SCoV 基因组的推定 pol 区域推导出来的。将合成重叠肽以覆盖整个pol la 和lb 开放阅读框。肽的长度为 20-60 个氨基酸。将通过 ELISA 方法筛选单个肽，以确定其与确诊 SARS 患者血清的反应性。肽上的血清学反应表位将被精炼以获得最佳序列和构象。新鉴定的非结构肽抗原将被添加到SARS ELISA的固相抗原制剂中。肽和试剂盒将按照 cGMP 标准进行制造和表征。大规模临床试验将在亚洲的设施中对大量 SARS 阳性血清进行。