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

基于量子点的疟疾定性和定量多重试纸测试 I

• 批准号: 8058384
• 负责人: RUOBING WANG
• 金额: $ 30万
• 依托单位: OCEAN NANOTECH, LLC
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8058384
• 关键词: Absorbent Pads; Affect; Agreement; Antibodies; Antigens; Biological; Biological Assay; Biological Markers; Blood Banks; Blood donor; Blood specimen; Caring; Cessation of life; Child; Color; Communicable Diseases; Country; Culicidae; Detection; Diagnosis; Diagnostic; Diagnostic tests; Disease; Drug resistance; Drug usage; Dyes; Exhibits; Fluorescence; Glutamate Dehydrogenase; Goals; Health Personnel; Human; Infection; Insecticide Resistance; Lactate Dehydrogenase; Lateral; Legal patent; Licensing; Life; Malaria; Measurement; Medicine; Membrane; Methods; Metric; Military Personnel; Oceans; Parasitemia; Parasites; Pathogen detection; Pharmaceutical Preparations; Phase; Plasmodium; Plasmodium falciparum; Property; Quantum Dots; Reader; Reagent Strips; Reliability of Results; Reproducibility; Sampling; Science; Side; System; Technology; Testing; Time; Travel; United States National Institutes of Health; Vaccines; Visual; Whole Blood; base; density; diagnostic accuracy; drug candidate; drug development; histidine-rich proteins; instrument; light intensity; multiplex detection; nanoparticle; novel; quantum; response; single molecule; vaccine candidate; vaccine development

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.

描述（由申请人提供）：我们的目标是开发易于使用、廉价和灵敏的基于量子点珠（QDB）的试纸条测试，用于人类疟疾感染的多重诊断，称为MAL-QDB测试”，其可以在<20分钟的测定中检测低至1个寄生虫/uL。MAL-QDBTest”包括1）用于定性目视检测的多重试纸检测（图1），2）试纸检测试剂，以及3）称为QD分析仪的可选手持式荧光阅读器”。MAL-QDB测试”在多重试纸测试中使用新型量子点珠，该测试用于a）使用目视检查的定性检测和B）使用QD-分析仪的定量检测”。MAL-QDTest”使用新型量子点珠（QDB），其可用于区分a）恶性疟原虫与间日疟原虫; B）检测引起疟疾的其他疟原虫; c）通过检测pGluDH区分活的与死的寄生虫;以及d）使用QD-分析仪定量存在的寄生虫的数量。它使用量子点珠（QDB），其是含有数百个量子点的纳米颗粒（NP），其表现出比有机染料更大的颜色稳定性和高光强度。具有几个原子到10，000个原子的量子点的量子限制特性可以实现单分子检测。与QD分析仪连接，基于QDB的多重检测将提供更高的灵敏度，更大的重现性，更低的检测限和更高的诊断准确性。 拟议的多重MAL-QDBTest”使用专利的QDB，可以与选择性和特异性抗体结合，用于捕获和检测疟疾感染的抗原生物标志物。迄今为止，还没有使用QDB的现有疟疾测定法，QDB是数百个NP的囊，其表现出比具有检测单个分子潜力的有机染料大100%的颜色稳定性和高光强度。 非常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灵敏度和可靠性的影响，而且不能检测到低于100个寄生虫/uL可能是致命的。目前的快速诊断测试的灵敏度可能>85%，但仅在200个寄生虫/uL或更高。这些不能用于需要1个寄生虫/uL检测的药物/疫苗开发。 拟议的MAL-QDB测试”预计将减轻每年超过5亿例疟疾病例的诊断，其中有1-3百万人死亡，以及药物和疫苗开发障碍。目前，没有针对这种疾病的许可疫苗。在美国，由于前往疟疾流行国家的旅行增加以及在全球部署超过30万名美国军事人员，疟疾可能重新出现。此外，由于最近前往流行国家而被拒绝献血的人数超过10%，这可能导致全国各地血库短缺。因此，需要开发与QD分析仪接口的所提出的MAL-QDBTest，用于快速、易于使用、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。 公共卫生关系：所提出的用于人类疟疾感染的多重诊断的基于量子点的试纸测试预计将减轻诊断以及药物和疫苗障碍，这些障碍每年影响> 5亿例疟疾病例和1至300万例死亡，其中大多数是幼儿。虽然最近在医学、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，这种疾病是对部署在世界各地的美军的主要传染病威胁。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的献血者人数超过10%，可能导致全国各地血库短缺。