Rapid LFIA blood test for SCD using highly specific monoclonal antibodies

项目摘要/摘要 本提案描述了快速免疫检测试剂盒开发的第二阶段。 血液中血红蛋白变种的测定，作为一种护理方法 贫瘠地区镰状细胞病及其携带者的诊断试验 全球范围内的设置。今天，SCD和SCD性状的确定需要漫长和 昂贵的实验室分析在低资源环境下不可用，以及&gt；80% 有SCD和SCD特征的人生活在提供医疗保健的地区， 设置。尽管如此，新生儿筛查计划已经导致了显著的 早期治疗可改善患者预后，预防并发症 疾病。 拟议的检测试剂盒将是一种横向流动免疫分析，原则上类似于家庭 验孕测试。该测试设计是基于针对 SCD相关血红蛋白变异体之间的单一氨基酸差异。在第一阶段， 申请人开发了一种工作原型血型检测试剂盒，该试剂盒被证明是 正确识别99%的检测全血样本的表型，包括每个 相关的SCD和携带者表型。原型测试需要1微升的 血液和结果的时间不到20分钟。在第二阶段，检测试剂盒将进一步 经过改进和简化，将开发出完整的可制造产品。 将进行临床验证研究，以确定 该产品，包括在巴西低资源环境下进行的研究。在 成功完成第二阶段，检测试剂盒将准备好制造和 提交世界各地的监管机构审批。项目的预计成本 血型检测试剂盒的价格将为2.00美元，这一价格点将允许广泛接受 这一检测试剂盒在发展中国家。

PROJECT SUMMARY/ABSTRACT This proposal describes Phase II of the development of a rapid immunoassay test kit for the determination of hemoglobin variants in blood, to be used as a point-of-care diagnostic test for sickle cell disease (SCD) and SCD carrier trait in low-resource settings worldwide. Today, determination of SCD and SCD trait requires lengthy and expensive laboratory analyses that are not available in low-resource settings, and >80% of individuals with SCD and SCD trait live in areas where healthcare is provided in those settings. Nevertheless, newborn screening programs have resulted in significant improvement of patient outcomes due to early treatment to prevent complications of the disease. The proposed test kit will be a lateral flow immunoassay, similar in principle to a home pregnancy test. The test design is based on monoclonal antibodies (MAbs) specific for the single-amino-acid differences between SCD-related hemoglobin variants. In Phase I, the applicant developed a working prototype hemotyping test kit which was shown to correctly identify the phenotype of >99% of tested whole blood samples, including each relevant SCD and carrier phenotype. The prototype test requires a 1-microliter volume of blood and the time-to-result is under 20 minutes. In Phase II, the test kit will be further improved and simplified, and a complete manufacturable product will be developed. Clinical validation studies will be carried out to determine the real-world performance of the product, including studies performed in low-resource settings in Brazil. At the successful conclusion of Phase II, the test kit will be ready for manufacturing and submission for approval to regulatory agencies worldwide. The estimated cost of the hemotyping test kit will be $2.00, a price point that will allow widespread acceptance of this test kit in developing countries.