Quantum Dot-based Qualitative and Quantitative Multiplex Strip Test for Malaria I

描述(由申请人提供)：我们的目标是开发易于使用，廉价，和敏感的量子点珠(QDB)为基础的条带测试的多重诊断人类疟疾感染称为MAL-QDB测试“，可以检测到低至1寄生虫/ul在一个&lt；20分钟的测试。MAL-QDBTest“包括1)用于定性视觉检测的多重试纸测试(图1)，2)用于试纸测试的试剂，以及3)一个可选的被称为QD-Analyzer的手持荧光读取器”。MAL-QDBTest“在多重试条测试中使用新的量子点珠子，用于a)使用目测进行定性检测，b)使用QD-Analyzer进行定量检测”。MAL-QDTest“使用新的量子点珠(QDB)，可用于区分a)恶性疟原虫和Pvivax；b)检测其他导致疟疾的疟原虫；c)通过检测pGluDH区分活寄生虫和死亡寄生虫；以及d)使用QD-Analyzer量化存在的寄生虫数量。它使用量子点珠(QDB)，这是一种纳米粒子(NP)，包含数百个量子点，表现出比有机染料更稳定的颜色和更高的光强度。几个原子到一万个原子的量子点的量子受限特性可以实现单分子检测。与QD-Analyzer相结合，基于QDB的多路检测将提供更高的灵敏度、更大的重复性、更低的检测极限和更高的诊断准确性。拟议的多重MAL-QDBTest“使用可与选择性和特定抗体结合的专利QDB，用于捕获和检测疟疾感染的抗原生物标记物。到目前为止，还没有现有的疟疾检测方法使用QDB，QDB是数百种具有颜色稳定性和高光强的NPs的囊状结构，它们比具有检测单个分子潜力的有机染料大100%。迫切需要开发更好的疟疾寄生虫检测方法，因为目前的诊断不仅受到结果的敏感性和可靠性的影响，而且无法检测到低于100个寄生虫/u1的寄生虫可能是致命的。目前快速诊断试验的敏感性可能为85%，但只能在200个寄生虫/微升或更高的水平上。这些不能用于需要1个寄生虫/微升检测的药物/疫苗开发。拟议的MAL-QDBTest“预计将减少每年5亿疟疾病例的诊断，其中有100-300万人死亡，以及药物和疫苗开发的障碍。目前，还没有获得许可的针对这种疾病的疫苗。在美国，由于前往流行国家的旅行增加，以及在世界各地部署30万美国军事人员，疟疾可能会重新出现。此外，由于最近前往流行国家而被拒绝献血的人数一直在10%，这可能导致全国各地血库短缺。因此，有必要开发拟议的MAL-QDBTest“与QD-Analyzer接口，以实现快速、易用、可靠、定量和廉价的疟疾诊断方法，并快速验证候选药物和疫苗。 与公共卫生相关：拟议的基于量子点的条带测试用于人类疟疾感染的多重诊断，预计将减少诊断以及药物和疫苗障碍，这些障碍每年影响5亿疟疾病例和100万至300万死亡病例，其中大部分是幼儿。尽管最近在医学、科学和技术方面取得了进展，但还没有针对这种疾病的许可疫苗，这种疾病是部署在世界各地的美军面临的主要传染病威胁。在美国，由于抗药性寄生虫和抗药性蚊子的传播，疟疾最近被认为有很高的重新出现的可能性，因此，由于最近前往流行国家而被拒绝献血的人数一直超过10%，可能导致全国各地血库出现短缺。

DESCRIPTION (provided by applicant): Our goal is to develop easy to use, inexpensive, and sensitive quantum dots beads (QDBs) based strip test for multiplex diagnosis of human malaria infection called MAL-QDBTest" that can detect as low as 1 parasite/uL in a <20 min assay. The MAL-QDBTest" consists of 1) multiplexed strip test for qualitative visual detection (Fig. 1), 2) reagents for the strip test, and 3) an optional a handheld fluorescence reader called QD-Analyzer". The MAL-QDBTest" uses novel quantum dots beads in a multiplex strip test that is used both for a) qualitative detection using visual inspection and b) quantitative detection using the QD- Analyzer". The MAL-QDTest" uses novel quantum dots beads (QDBs) that can be used to differentiate a) Pfalciparum from Pvivax; b) detect other Plasmodia causing malaria; c) differentiate live from dead parasites through the detection of pGluDH; and d) quantify the number of parasites present with the use of the QD- Analyzer. It uses quantum dots beads (QDBs) which are nanoparticles (NPs) containing hundreds of quantum dots that exhibit color stability and high light intensity greater than organic dyes. Quantum confined properties of quantum dots with a few atoms to 10,000 atoms can enable single molecule detection. Interfaced with the QD-Analyzer, the QDB-based multiplex detection will provide higher sensitivity, greater reproducibility, lower detection limits, and increased diagnostic accuracy. The proposed multiplex MAL-QDBTest" uses patented QDBs that can be conjugated to selective and specific antibodies for the capture and detection of antigen biomarkers of malaria infection. To date, there is no existing malaria assay that uses QDBs which are sacs of hundreds of NPs that exhibit color stability and high light intensity that are 100% greater than organic dyes with the potential to detect a single molecule. There is a great need for developing better detection methods for malaria parasites because not only does current diagnosis suffer from sensitivity and reliability of results, these cannot detect lower than 100 parasites /uL may be fatal. The sensitivity of current rapid diagnostic tests may be >85% but only at 200 parasites/uL or higher. These cannot be used for drug/vaccine development that requires 1 parasite/uL detection. The proposed MAL-QDBTest" is anticipated to alleviate the diagnosis of > 500 million cases of malaria each year with 1-3 million deaths and the drug and vaccine development hurdles. Currently, there is no no licensed vaccine against this disease. In the US, malaria may re-emerge due to increased travel to endemic countries as well as the deployment of the >300,000 US military personnel worldwide. Furthermore, the number of blood donors who had been refused as a result of recent travel to endemic countries has been > 10% that may have caused shortage at blood banks around the country. Thus, there is a need to develop the proposed MAL-QDBTest" interfaced with the QD-Analyzer for fast, easy to use, reliable, quantitative, and inexpensive methods for malaria diagnosis and to quickly validate drug and vaccine candidates. PUBLIC HEALTH RELEVANCE: The proposed quantum dots based strip test for multiplex diagnosis of human malaria infection is anticipated to alleviate the diagnostics as well as the drug and vaccine hurdles that affect > 500 million cases of malaria each year and 1 to 3 million deaths, majority of whom are young children. Although there are recent advances in medicine, science, and technology, there is no licensed vaccine against this disease that is a major infectious disease threat to U.S. forces deployed worldwide. In the US, malaria is recently considered to have a high potential to re-emerge due to the spread of drug resistant parasites and insecticide-resistant mosquitoes and as a result the number of blood donors who had been refused as a result of recent travel to endemic countries has been more than 10% possibly causing shortage at blood banks around the country.