Project-004

项目4概要 以青蒿素为基础的联合疗法（ACT）作为简单疟原虫的一线治疗 恶性疟疾在减少全球疟疾负担方面发挥了不可或缺的作用。因此 伪造和不合标准的青蒿素综合疗法的流通已成为一个新出现的全球危机， 在疟疾控制方面取得的成就。然而，这一问题的严重程度尚不清楚，迫切需要 对伪造的青蒿素综合疗法进行系统监测。由于青蒿素综合疗法的传统质量控制方法大多 需要昂贵的仪器和大量的技术支持，即时（POC）测试的开发 用于在地方病环境下检测青蒿素综合疗法中的活性成分是非常需要的。另外作为 许多疟疾流行的国家正在朝着消灭疟疾、大规模药物管理（MDA）和 季节性疟疾化学预防（SMC）计划正在越来越多地部署。一种廉价且高通量的检测方法可用于测量长效ACT伴侣药物的血清水平， 允许对MDA和SMC计划的覆盖率和遵守率进行成本效益评估。到 为了满足这些需求，我们开发了高灵敏度和可靠的酶联免疫吸附测定法 （ELISA）用于青蒿素及其衍生物的准确定量。我们进一步开发了一种基于抗体的 作为POC检测的试纸法，用于ACT中ART及其衍生物的定性和半定量评价 这些药物经过试点测试，快速、方便、易于在流行地区使用。基于 这些最近的成功，我们建议1）使用新的POC诊断工具，系统地调查 问题最严重的大湄公河次区域青蒿素假药的严重程度 严重，2）开发一系列特异性单克隆抗体的三个ACT合作伙伴药物使用， 免疫原的创新设计，以及3）开发和优化用于定量和 检测市售药物和人血浆中的ACT伴侣药物。我们预计ELISA 将为精确定量ACT中的活性药物成分提供非常方便的工具， 实验室，而组合试纸提供POC设备，用于偏远地方病患者的ACT质量控制。 设置.

PROJECT 4 SUMMARY Artemisinin-based combination therapies (ACTs) as the frontline treatment of uncomplicated Plasmodium falciparum malaria have played an indispensable role in reducing the global malaria burden. Thus, the circulation of falsified and substandard ACTs has become an emerging global crisis, which greatly threatens the gain made in malaria control. Yet, the extent of this problem is not known and there is a pressing need for systematic surveillance efforts of falsified ACTs. Since traditional methods for quality control of ACTs mostly require expensive instruments and substantial technical support, the development of a point-of-care (POC) test for the detection of the active ingredients in ACTs under endemic settings is highly desired. In addition, as many malaria-endemic nations are moving towards malaria elimination, mass drug administration (MDA) and seasonal malaria chemoprevention (SMC) programs are being increasingly deployed. A cheap and higher-throughput assay that could be used to measure the serum levels of the long-lasting ACT partner drugs will allow cost-effective assessment of the coverage and compliance rates in the MDA and SMC programs. To address these needs, we developed highly sensitive and reliable enzyme-linked immunosorbent assays (ELISAs) for accurate quantitation of artemisinin and its derivatives. We further developed an antibody-based dipstick assay as a POC test for qualitative and semi-quantitative evaluation of ART and its derivatives in ACT drugs, which have been pilot-tested to be fast, convenient, and easy to perform in endemic sites. Based on these recent successes, we propose to 1) use the new POC diagnostic tools to systematically investigate the extent of falsified artemisinin drugs in the Greater Mekong Subregion, where the problem had been most serious, 2) to develop a series of specific monoclonal antibodies for three ACT partner drugs using an innovative design of immunogens, and 3) develop and optimize ELISAs and dipsticks for quantification and detection of ACT partner drugs in both commercial drugs and human plasma. We anticipate that the ELISAs will provide highly convenient tools for accurate quantification of active pharmaceutical ingredients in ACTs in the laboratory, while the combination dipsticks offer POC devices for quality control of ACTs in remote endemic settings.