Rapid LFIA blood test for SCD using highly specific monoclonal antibodies

项目总结/摘要 该提案描述了快速免疫测定试剂盒的第二阶段开发， 确定血液中的血红蛋白变体，用作即时护理 低资源地区镰状细胞病（SCD）和SCD携带者性状的诊断试验 世界各地的设置。今天，SCD和SCD性状的确定需要长时间和长时间的研究。 昂贵的实验室分析，在低资源环境中不可用，并且> 80% 患有SCD和SCD特征的人居住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着的 由于早期治疗以预防并发症， 疾病 拟议的检测试剂盒将是一个侧流免疫测定，类似于家庭的原则， 验孕棒试验设计基于对以下物质具有特异性的单克隆抗体（MAb）： SCD相关血红蛋白变体之间的单个氨基酸差异。在第一阶段， 申请人开发了一种工作原型血型鉴定试剂盒， 正确识别> 99%的检测全血样本的表型，包括 相关SCD和携带者表型。原型测试需要1微升体积的 血液和时间的结果是在20分钟内。在第二阶段，检测试剂盒将进一步 改进和简化，并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中进行的研究。在 第二阶段的成功结束，检测试剂盒将准备生产， 提交给全球监管机构审批。的估计费用 血型检测试剂盒将是2.00美元，一个价格点，将允许广泛接受 在发展中国家使用这种检测试剂盒。

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.